CN1735605A - Phenylalanine derivative is used for the treatment of or prevent diabetes as depeptidyl peptidase inhibitors - Google Patents

Abstract

The present invention relates to phenylalanine derivative, this derivative is the inhibitor (" DP-IV inhibitor ") of dipeptidyl peptidase-IV enzyme, is used for the treatment of or prevents to relate to the disease of dipeptidyl peptidase-IV enzyme, for example diabetes, especially diabetes B.The present invention also relates to comprise the pharmaceutical composition of these compounds and these compounds and composition in prevention or treat purposes in the disease that relates to the dipeptidyl peptidase-IV enzyme.

Description

Phenylalanine derivatives as dipeptidyl peptidase inhibitors for the treatment or prevention of diabetes

Background of the invention

Diabetes mellitus refers to a disease process of various etiologies characterized by elevated plasma glucose levels or hyperglycemia in the fasted state or following administration of glucose during an oral glucose tolerance test. Persistent or uncontrolled hyperglycemia is associated with increased and earlier morbidity and mortality. In general, abnormalities in glucose homeostasis are directly and indirectly associated with altered lipid, lipoprotein, and apolipoprotein metabolism, as well as other metabolic and hemodynamic disorders. Thus, patients with type 2 diabetes are at greatly increased risk of macrovascular and microvascular complications, including coronary heart disease, stroke, peripheral vascular disease, hypertension, nephropathy, neuropathy, and retinopathy. Therefore, therapeutic control of glucose homeostasis, lipid metabolism and hypertension is crucial in the clinical management and treatment of diabetes.

There are usually two identifiable forms of diabetes. In type 1 diabetes, or insulin-dependent diabetes mellitus (IDDM), the patient produces little or no insulin, the hormone that regulates glucose utilization. In type 2 diabetes, or non-insulin-dependent diabetes mellitus (NIDDM), patients usually have the same or higher plasma insulin levels compared to non-diabetics; however, these patients have developed insulin-stimulated insulin-stimulating Resistance to glucose and lipid metabolism, the sensitive tissues being muscle, liver and adipose tissue, and even elevated plasma insulin levels are not sufficient to overcome marked insulin resistance.

Insulin resistance is not primarily due to a reduced number of insulin receptors, but rather an as-yet-understood post-binding defect of insulin receptors. This resistance to insulin response results in insufficient insulin-activated glucose uptake, oxidation, and storage in muscle, and inappropriate inhibition by insulin of lipolysis in adipose tissue and glucose production and secretion in the liver.

The available treatments for type 2 diabetes have remained largely unchanged over the years and have recognized limitations. Physical exercise and dietary calorie reduction unexpectedly improve diabetes symptoms, and adherence to this treatment is very limited because of a sedentary lifestyle and excessive food consumption (particularly those high in saturated fat) that are entrenched. By giving sulfonylureas (such as tolbutamide and glipizide) or meglitinide, which stimulate pancreatic beta cells to secrete more insulin, and/or by injection when sulfonylureas or meglitinide are ineffective Insulin, which increases plasma levels of insulin, can lead to insulin concentrations high enough to stimulate insulin-resistant tissues. However, administration of insulin or insulin secretagogues (sulfonylureas or meglitinide) can lead to dangerously low levels of plasma glucose and even higher levels of insulin resistance can occur due to increased plasma insulin levels. Biguanides increase insulin sensitivity, leading to correction of some hyperglycemia. However, two biguanides, phenformin and metformin, can induce lactic acidosis and nausea/diarrhea. Metformin has fewer side effects than phenformin and is often prescribed to treat type 2 diabetes.

The glitazones (ie, 5-benzylthiazolidine-2,4-dione) are a recently described class of compounds that have the potential to ameliorate many of the symptoms of type 2 diabetes. These drugs essentially increase insulin sensitivity in muscle, liver, and adipose tissue in certain animal models of type 2 diabetes, resulting in partial or complete correction of elevated plasma glucose levels without hypoglycemia. Glitazones currently on the market are agonists of peroxisome proliferator-activated receptors (PPARs), mainly the PPAR-γ subtype. PPAR-gamma agonism is generally believed to be responsible for the observed improvements in insulin sensitivity with glitazones. Newer PPAR agonists being tested for the treatment of type 2 diabetes are agonists of the alpha, gamma, or delta subtypes, or combinations thereof, and in many cases are chemically distinct from the glitazones (ie, they are not thiazolidinediones ). Some glitazones, such as troglitazone, develop serious side effects (eg, hepatotoxicity).

Other methods of treating this disease are still under investigation. Neochemical approaches that have been recently introduced or are still under development include treatment with alpha-glucosidase inhibitors (eg, acarbose) and protein tyrosine phosphatase-1B (PTP-1B) inhibitors.

Compounds that are dipeptidyl peptidase-IV ("DP-IV" or "DPP-IV") enzyme inhibitors are also under investigation as drugs useful in the treatment of diabetes, particularly type 2 diabetes. See for example WO 97/40832, WO 98/19998, U.S. Patent No. 5939560, Bioorg.Med.Chem.Lett . , 6:1163-1166 (1996) and Bioorg.Med.Chem.Lett. , 6:2745-2748 ( 1996). The use of DP-IV inhibitors in the treatment of type 2 diabetes is based on the fact that DP-IV tends to inactivate glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP) in vivo. GLP-1 and GIP are incretins that are produced when food is consumed. Incretins stimulate insulin production. Inhibition of DP-IV leads to reduced inactivation of incretins, which in turn leads to increased effectiveness of incretins in stimulating pancreatic insulin production. DP-IV inhibition thus results in elevated plasma insulin levels. Advantageously, because the body produces incretins only when food is consumed, DP-IV inhibition is not expected to increase insulin levels at inappropriate times, such as between meals, which could lead to excessive hypoglycemia. Inhibition of DP-IV action is therefore expected to increase insulin without increasing the risk of hypoglycemia, a deleterious side effect associated with the use of insulin secretagogues.

DP-IV inhibitors also have other therapeutic uses discussed herein. DP-IV inhibitors have not been intensively studied to date, especially for uses other than diabetes. New compounds are needed to discover improved DP-IV inhibitors for the treatment of diabetes and other underlying diseases and conditions.

Summary of the invention

The present invention relates to phenylalanine derivatives which are dipeptidyl peptidase-IV enzyme inhibitors ("DP-IV inhibitors") for use in the treatment or prevention of diseases involving dipeptidyl peptidase-IV enzymes, such as Diabetes, especially type 2 diabetes. The invention also relates to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions in the prevention or treatment of diseases involving dipeptidyl peptidase-IV enzymes.

Detailed description of the invention

The present invention relates to phenylalanine derivatives useful as inhibitors of dipeptidyl peptidase-IV. Compounds of the present invention are described by the following structural formula I or a pharmaceutically acceptable salt thereof:

in:

Each n is independently 0, 1 or 2;

m and p are independently 0 or 1;

X is _CH2 , S, CHF or _CF2 ;

W and Z are each independently CH ₂ , CHF or CF ₂ ;

R ¹ is hydrogen or cyano;

Each R ^is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, cyano, trifluoromethyl, trifluoromethoxy and hydroxy;

^R is aryl, heteroaryl or heterocyclyl, wherein aryl, heteroaryl and heterocyclyl are unsubstituted or substituted with ^1-5 R substituents;

^R2 is selected from

hydrogen,

C _1-10 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen or hydroxyl,

C _2-10 alkenyl, wherein alkenyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen or hydroxyl,

(CH ₂ ) _n -aryl, wherein aryl is unsubstituted or with 1-5 independently selected from hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1- Substituents of ₆ alkoxy groups, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens,

(CH ₂ ) _n -heteroaryl, wherein the heteroaryl is unsubstituted or with 1-3 independently selected from hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C Substituents of _1-6 alkoxy groups, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n -heterocyclic group, wherein the heterocyclic group is unsubstituted or with 1-3 independently selected from oxo, hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkane Substituents of radicals and C _1-6 alkoxy groups, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens,

(CH ₂ ) _n -C _3-6 cycloalkyl, wherein cycloalkyl is unsubstituted or with 1-3 independently selected from halogen, hydroxyl, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 Substituents of alkyl and C _1-6 alkoxy, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n COOH,

(CH ₂ ) _n COOC _1-6 alkyl,

(CH ₂ ) _n CONR ⁶ R ⁷ , wherein R ⁶ and R ⁷ are independently selected from hydrogen, tetrazolyl, thiazolyl, (CH ₂ ) _n -phenyl, (CH ₂ ) _n -C _3-6 cycloalkyl and C _1-6 alkyl, wherein alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen and hydroxy, and wherein phenyl and cycloalkyl are unsubstituted or substituted with 1-5 independently selected from halogen , hydroxy, C _1-6 alkyl and C _1-6 alkoxy substituents, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogens;

Or wherein R ⁶ and R ⁷ form a heterocycle selected from azetidine, pyrrolidine, piperidine, piperazine and morpholine together with the nitrogen atom to which they are attached, wherein the heterocycle is unsubstituted or replaced with 1-3 Substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy are substituted, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen;

wherein any methylene (CH ₂ ) carbon atom in R ² is unsubstituted or with 1-2 independently selected from halogen, hydroxyl and C _1-4 alkyl unsubstituted or substituted with 1-5 halogen group replacement;

Each R ⁵ is independently selected from

halogen,

cyano,

Oxo,

hydroxyl,

C _1-6 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 halogens,

C _1-6 alkoxy, wherein alkoxy is unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n -NR ⁶ R ⁷ ,

(CH ₂ ) _n -CONR ⁶ R ⁷ ,

(CH ₂ ) _n -OCONR ⁶ R ⁷ ,

(CH ₂ ) _n -SO ₂ NR ⁶ R ⁷ ,

(CH ₂ ) _n -SO ₂ R ⁹ ,

(CH ₂ ) _n -NR ⁸ SO ₂ R ⁹ ,

(CH ₂ ) _n -NR ⁸ CONR ⁶ R ⁷ ,

(CH ₂ ) _n -NR ⁸ COR ⁸ ,

(CH ₂ ) _n -NR ⁸ CO ₂ R ⁹ ,

( _CH2 ) _n -COOH,

(CH ₂ ) _n -COOC _1-6 alkyl,

(CH ₂ ) _n -aryl, wherein aryl is unsubstituted or is independently selected from halogen, hydroxyl, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl, C _3- Substituents of ₆ cycloalkyl and C _1-6 alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n -heteroaryl, wherein heteroaryl is unsubstituted or with 1-3 independently selected from hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl, C Substituents of _3-6 cycloalkyl and C _1-6 alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n -heterocyclic group, wherein the heterocyclic group is unsubstituted or with 1-3 independently selected from oxo, hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkane Substituents of radicals, C _3-6 cycloalkyl and C _1-6 alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogens,

(CH ₂ ) _n -C _3-6 cycloalkyl, wherein cycloalkyl is unsubstituted or with 1-3 substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy Substituted, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

wherein any methylene (CH ₂ ) carbon atom in R ⁵ is unsubstituted or with 1-2 independently selected from halogen, hydroxyl and C _1-4 alkyl unsubstituted or substituted with 1-5 halogen group replacement;

Each R ⁹ is independently selected from tetrazolyl, thiazolyl, (CH ₂ ) _n -phenyl, (CH ₂ ) _n -C _3-6 cycloalkyl and C _1-6 alkyl, wherein the alkyl is unsubstituted or Substituted with 1-5 halogen, and wherein phenyl and cycloalkyl are unsubstituted or substituted with 1-5 substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens, and wherein any methylene (CH ₂ ) carbon atom in R ⁸ is unsubstituted or substituted with 1-2 independently selected from halogen, hydroxyl and Unsubstituted or substituted with 1-5 halogen-substituted C _1-4 alkyl groups;

Each R ⁸ is hydrogen or R ⁹ .

In one embodiment of the compounds of the invention, the carbon atom marked with ^* has the stereochemical configuration as described in formula Ia:

Wherein R ³ is hydrogen or fluorine;

W, X, Z, m, p, R ¹ , R ² and R ⁴ are as defined above.

In this embodiment of a class of compounds of the invention, the carbon atom attached to ^R , marked with ^** , has the stereochemical configuration as described in Formula Ib:

wherein ^R3 is hydrogen or fluorine,

W, X, Z, m, p, R ¹ , R ² and R ⁴ are as defined above.

In a second embodiment of the compounds of the invention, as described in formula Ic, m is 1 and p is 0:

wherein ^R3 is hydrogen or fluorine,

W, X, R ¹ , R ² and R ⁴ are as defined above.

One class of this embodiment includes compounds wherein the carbon atom marked with ^* and the carbon atom marked with ^** have the stereochemical configuration as described in Formula Id:

wherein R ³ is hydrogen or fluorine, W, X, R ¹ , R ² and R ⁴ are as defined above.

In a subclass of such compounds of the invention, ^R1 is hydrogen, W is _CH2 , and X is _CH2 , CHF or _CF2 .

In a third embodiment of the compounds of the invention, as described in formula Ie, R ^is hydrogen, X is CHF, m and p are 0:

Wherein R ³ is hydrogen or fluorine, R ² and R ⁴ are as defined above.

One class of this embodiment includes compounds wherein the carbon atom marked with ^* has the stereochemical configuration as described in Formula If:

Wherein R ³ is hydrogen or fluorine, R ² and R ⁴ are as defined above.

In a fourth ^embodiment of the compounds of the invention, as depicted in formula Ig, R is hydrogen, m and p are 1:

Wherein R ³ is hydrogen or fluorine;

W, X, Z, ^R2 and ^R4 are as defined above.

One class of this embodiment includes compounds wherein the carbon atom marked with ^* has the stereochemical configuration as described in Formula Ih:

wherein ^R3 is hydrogen or fluorine, W, X, Z, ^R2 and ^R4 are as defined above.

In a subclass of this class, W and Z are _CH2 and X is CHF or _CF2 .

In a fifth embodiment of the compounds of this invention, R ^is selected from

C _1-6 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen or hydroxyl,

C _2-6 alkenyl, wherein alkenyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen or hydroxyl,

(CH ₂ ) _n -C _3-6 cycloalkyl, wherein cycloalkyl is unsubstituted or with 1-3 independently selected from halogen, hydroxyl, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 Substituents of alkyl and C _1-6 alkoxy, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n COOH,

(CH ₂ ) _n COOC _1-6 alkyl,

(CH ₂ ) _n CONR ⁶ R ⁷ , wherein R ⁶ and R ⁷ are independently selected from hydrogen, tetrazolyl, thiazolyl, (CH ₂ ) _n -phenyl, (CH ₂ ) _n -C _3-6 cycloalkyl and C _1-6 alkyl, wherein alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen and hydroxy, and wherein phenyl and cycloalkyl are unsubstituted or substituted with 1-5 independently selected from halogen , hydroxy, C _1-6 alkyl and C _1-6 alkoxy substituents, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogens;

Or wherein R ⁶ and R ⁷ form a heterocycle selected from azetidine, pyrrolidine, piperidine, piperazine and morpholine together with the nitrogen atom to which they are attached, wherein the heterocycle is unsubstituted or replaced with 1-5 Substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy are substituted, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen;

wherein any methylene (CH ₂ ) carbon atom in R ² is unsubstituted or with 1-2 independently selected from halogen, hydroxyl and C _1-4 alkyl unsubstituted or substituted with 1-5 halogen group replaced.

In this embodiment of a class of compounds of the invention, R ^is selected from

C _1-3 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen or hydroxyl,

CH ₂ -C _3-6 cycloalkyl,

COOH,

COOC _1-6 alkyl,

CONR ⁶ R ⁷ , wherein R ⁶ and R ⁷ are independently selected from hydrogen, tetrazolyl, thiazolyl, (CH ₂ ) _n -phenyl, (CH ₂ ) _n -C _3-6 cycloalkyl and C _1-6 Alkyl, wherein alkyl is unsubstituted or substituted with 1-5 substituents independently selected from halogen and hydroxy, and wherein _phenyl and cycloalkyl are unsubstituted or substituted with 1-5 independently selected from halogen, hydroxy, C Substituents of _-6 alkyl and C _1-6 alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogens;

Or wherein R ⁶ and R ⁷ form a heterocycle selected from azetidine, pyrrolidine, piperidine, piperazine and morpholine together with the nitrogen atom to which they are attached, wherein the heterocycle is unsubstituted or replaced with 1-5 Substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy are substituted, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen;

A sixth embodiment of the invention includes compounds of formula Ii:

Wherein X is CH ₂ , S, CHF or CF ₂ ;

W and Z are each independently _CH2 , CHF or _CF2 ;

R ⁴ is phenyl, heteroaryl or heterocyclic group, wherein phenyl, heteroaryl and heterocyclic group are unsubstituted or substituted with ^1-3 R substituents;

^R2 is selected from:

methyl,

ethyl,

CH ₂ -cyclopropyl,

COOH,

COOMe,

COOEt,

CONHMe,

CONMe ₂ ,

_CONH2 ,

CONHEt,

CONMeCH ₂ Ph,

pyrrolidin-1-ylcarbonyl,

Azetidin-1-ylcarbonyl,

3-fluoroazetidin-1-ylcarbonyl,

Morpholin-4-ylcarbonyl,

[(Tetrazol-5-yl)amino]carbonyl;

each R is ^{independently} selected from:

halogen,

cyano,

Oxo,

hydroxyl,

C _1-6 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 halogens,

C _1-6 alkoxy, wherein alkoxy is unsubstituted or substituted with 1-5 halogen,

NR ⁶ R ⁷ ,

CONR ⁶ R ⁷ ,

OCONR ⁶ R ⁷ ,

SO ₂ NR ⁶ R ⁷ ,

SO ₂ R ₉ ,

NR ⁸ SO ₂ R ₉ ,

NR ⁸ CONR ⁶ R ⁷ ,

NR ⁸ COR ⁸ ,

NR ⁸ CO ₂ R ⁹ ,

COOH,

COOC _1-6 alkyl,

Aryl, wherein aryl is unsubstituted or substituted with 1-5 independently selected from halogen, hydroxyl, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1-6 alkoxy Substituted by radical, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

Heteroaryl, wherein heteroaryl is unsubstituted or with 1-3 independently selected from hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1-6 alkoxy Substituted by substituents, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens,

Heterocyclic group, wherein the heterocyclic group is unsubstituted or with 1-3 independently selected from oxo, hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1-6 Substituents of alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogen,

(CH ₂ ) _n -C _3-6 cycloalkyl, wherein cycloalkyl is unsubstituted or with 1-3 substituents independently selected from halogen, hydroxyl, C _1-6 alkyl and C _1-6 alkoxy Substituted, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens.

In a class of such embodiments, each R ^is independently selected from:

halogen,

cyano,

Oxo,

C _1-6 alkyl, wherein the alkyl is unsubstituted or substituted with 1-5 halogens,

C _1-6 alkoxy, wherein alkoxy is unsubstituted or substituted with 1-5 halogen,

CONR ⁶ R ⁷ ,

NR ⁸ COR ⁸ ,

SO ₂ R ⁹ ,

NR ⁸ SO ₂ R ⁹ ,

COOH,

COOC _1-6 alkyl,

Heteroaryl, wherein heteroaryl is unsubstituted or with 1-3 independently selected from hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1-6 alkoxy Substituted by substituents, wherein the alkyl and alkoxy groups are unsubstituted or substituted with 1-5 halogens,

Heterocyclic group, wherein the heterocyclic group is unsubstituted or with 1-3 independently selected from oxo, hydroxyl, halogen, CO ₂ H, C _1-6 alkoxycarbonyl, C _1-6 alkyl and C _1-6 Substituents of alkoxy, wherein the alkyl and alkoxy are unsubstituted or substituted with 1-5 halogens.

In a subclass of this class, ^R4 is selected from:

4-fluorophenyl,

2,4-difluorophenyl,

3,4-difluorophenyl,

2-chlorophenyl,

2-fluorophenyl,

3-(methylsulfonyl)phenyl,

3-(ethoxycarbonyl)phenyl,

3-carboxyphenyl,

3-(Aminocarbonyl)phenyl,

3-[(tert-butylamino)carbonyl]phenyl,

3-[(phenylamino)carbonyl]phenyl,

3-[(thiazol-2-ylamino)carbonyl]phenyl,

3-[(tetrazol-5-ylamino)carbonyl]phenyl,

3-[[(trifluoromethyl)sulfonyl]amino]phenyl,

3-(tetrazol-5-yl)phenyl,

4-fluoro-3-(tetrazol-5-yl)phenyl,

2-fluoro-5-(tetrazol-5-yl)phenyl,

3-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl,

4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl,

3-(5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)phenyl,

3-(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)phenyl,

3-(1,3,4-oxadiazol-2-yl)phenyl,

3-(1,2,4-triazol-3-yl)phenyl,

3-[5-(trifluoromethyl)-1,2,4-triazol-3-yl]phenyl,

3-(5-ethoxy-1,2,4-triazol-3-yl)phenyl,

pyridin-3-yl,

6-fluoro-pyridin-3-yl,

6-methoxypyridin-3-yl,

6-oxo-1,6-dihydropyridin-3-yl,

1-methyl-6-oxo-1,6-dihydropyridin-3-yl,

1-ethyl-6-oxo-1,6-dihydropyridin-3-yl,

5-bromo-1-methyl-6-oxo-1,6-dihydropyridin-3-yl,

imidazo[1,2-α]pyridin-6-yl,

[1,2,4]triazolo[4,3-α]pyridin-6-yl,

3-(trifluoromethyl)[1,2,4]triazolo[4,3-α]pyridin-6-yl,

3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-α]pyridin-6-yl,

2-Methyl-3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-α]pyridin-6-yl,

4-aminoquinazolin-6-yl,

2-(acetylamino)imidazo[1,2-α]pyridin-6-yl,

3-aminoimidazo[1,2-α]pyridin-6-yl,

3-carboxypyrazolo[1,5-α]pyridin-5-yl,

5-bromo-6-oxo-1,6-dihydropyridin-3-yl,

[1,2,4]triazolo[1,5-α]pyridin-6-yl,

[1,2,4]triazolo[1,5-α]pyridin-7-yl,

Pyrazolo[1,5-α]pyrimidin-5-yl.

Another class of this embodiment includes compounds of structure Ij:

^R2 is selected from:

methyl,

ethyl,

CH ₂ -cyclopropyl,

COOH,

COOMe,

COOEt,

CONHMe,

CONMe ₂ ,

_CONH2 ,

CONHEt,

CONMeCH ₂ Ph,

pyrrolidin-1-ylcarbonyl,

Azetidin-1-ylcarbonyl,

3-fluoroazetidin-1-ylcarbonyl,

Morpholin-4-ylcarbonyl,

[(Tetrazol-5-yl)amino]carbonyl;

^R4 is selected from:

4-fluorophenyl,

2,4-difluorophenyl,

3,4-difluorophenyl,

2-chlorophenyl,

2-fluorophenyl,

3-(methylsulfonyl)phenyl,

3-(ethoxycarbonyl)phenyl,

3-carboxyphenyl,

3-(Aminocarbonyl)phenyl,

3-[(tert-butylamino)carbonyl]phenyl,

3-[(phenylamino)carbonyl]phenyl,

3-[(thiazol-2-ylamino)carbonyl]phenyl,

3-[(tetrazol-5-ylamino)carbonyl]phenyl,

3-[[(trifluoromethyl)sulfonyl]amino]phenyl,

3-(tetrazol-5-yl)phenyl,

4-fluoro-3-(tetrazol-5-yl)phenyl,

2-fluoro-5-(tetrazol-5-yl)phenyl,

3-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl,

4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl,

3-(5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)phenyl,

3-(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)phenyl,

3-(1,3,4-oxadiazol-2-yl)phenyl,

3-(1,2,4-triazol-3-yl)phenyl,

3-[5-(trifluoromethyl)-1,2,4-triazol-3-yl]phenyl,

3-(5-ethoxy-1,2,4-triazol-3-yl)phenyl,

pyridin-3-yl,

6-fluoro-pyridin-3-yl,

6-methoxypyridin-3-yl,

6-oxo-1,6-dihydropyridin-3-yl,

1-methyl-6-oxo-1,6-dihydropyridin-3-yl,

1-ethyl-6-oxo-1,6-dihydropyridin-3-yl,

5-bromo-1-methyl-6-oxo-1,6-dihydropyridin-3-yl,

imidazo[1,2-α]pyridin-6-yl,

[1,2,4]triazolo[4,3-α]pyridin-6-yl,

3-(trifluoromethyl)[1,2,4]triazolo[4,3-α]pyridin-6-yl,

3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-α]pyridin-6-yl,

2-Methyl-3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-α]pyridin-6-yl,

4-aminoquinazolin-6-yl,

2-(acetylamino)imidazo[1,2-α]pyridin-6-yl,

3-aminoimidazo[1,2-α]pyridin-6-yl,

3-carboxypyrazolo[1,5-α]pyridin-5-yl,

5-bromo-6-oxo-1,6-dihydropyridin-3-yl,

[1,2,4]triazolo[1,5-α]pyridin-6-yl,

[1,2,4]triazolo[1,5-α]pyridin-7-yl,

Pyrazolo[1,5-α]pyrimidin-5-yl.

In a subclass of this class, W is _CH2 and X is CHF or _CF2 .

Illustrative, non-limiting examples of compounds of the invention useful as inhibitors of dipeptidyl peptidase-IV are the following compounds, or pharmaceutically acceptable salts thereof:

The following terms as used herein may be used.

"Alkyl" and other groups having the prefix "alk" such as alkoxy and alkanoyl refer to carbon chains and combinations thereof which may be linear or branched, unless the carbon chain is defined otherwise. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like. The term alkyl also includes cycloalkyl, and combinations of linear or branched alkyl chains and cycloalkyl structures when the specific number of carbon atoms permits, eg, _C3-10 . When the number of carbon atoms is not specified, the meaning is C _1-6 .

"Cycloalkyl" is a subset of alkyl and refers to a saturated carbocyclic ring having the specified number of carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Unless otherwise specified, cycloalkyl groups are generally monocyclic. Unless defined otherwise, cycloalkyl groups are saturated.

The term "alkoxy" refers to a straight or branched chain alkoxylate of the indicated number of carbon atoms (eg, C _1-6 alkoxy), or any number of carbon atoms within the range. [i.e. methoxy (MeO-), ethoxy, isopropoxy, etc.].

The term "alkylthio" refers to a straight or branched chain alkyl sulfide with the indicated number of carbon atoms (e.g. _C1-6 alkylthio), or a straight or branched chain alkyl group with any number of carbon atoms within the range Sulfides [i.e. methylthio (MeS-), ethylthio, isopropylthio, etc.].

The term "alkylamino" refers to a straight or branched chain alkylamine (eg, _C1-6 alkylamine) with the indicated number of carbon atoms, or any number of carbon atoms within the range. [i.e. methylamino, ethylamino, isopropylamino, tert-butylamino, etc.].

The term "alkylsulfonyl" refers to a straight or branched chain alkyl sulfone with the indicated number of carbon atoms (e.g. _C1-6 alkylsulfonyl), or a straight or branched chain alkane with any number of carbon atoms within the range. Sulfone [that is, methylsulfonyl (MeSO ₂ -), ethylsulfonyl, isopropylsulfonyl, etc.].

The term "alkoxycarbonyl" refers to the straight or branched chain esters of the carboxylic acid derivatives of the present invention (e.g., C _1-6 alkoxycarbonyl) with the indicated number of carbon atoms, or any number of carbon atoms within the range. Straight chain or branched esters of carboxylic acid derivatives [ie methoxycarbonyl (MeOCO-), ethoxycarbonyl or butoxycarbonyl] are invented.

"Aryl" means a monocyclic or polycyclic aromatic ring system comprising carbon ring atoms. Preferred aryl groups are monocyclic or bicyclic 6-10 membered aromatic ring systems. Phenyl and naphthyl are preferred aryl groups. The most preferred aryl is phenyl.

"Heterocycle" and "heterocyclyl" refer to a saturated or unsaturated non-aromatic ring or ring system comprising at least one heteroatom selected from O, S and N, and also include the oxidized forms of sulfur, namely SO and _SO2 . Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine, 1,3-dioxolane, imidazolidine , imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane, 1,3-dithiane, oxysulfur Heterocyclohexane, thiomorpholine, etc.

"Heteroaryl" refers to an aromatic or partially aromatic heterocyclic ring comprising at least one ring heteroatom selected from O, S, and N. Heteroaryl also includes heteroaryl fused to other kinds of rings such as aryl, cycloalkyl, and non-aromatic heterocyclyl. Examples of heteroaryl groups include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, 2-oxo-(1H)-pyridyl (2-hydroxy-pyridyl), oxazolyl, 1 , 2,4-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furyl, triazinyl, thienyl, Pyrimidinyl, pyrazinyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, dihydrobenzofuranyl, indolinyl, pyridazinyl, indole Azolyl, isoindolyl, dihydrobenzothienyl, indolizinyl, cinnolinyl, 2,3-diazinyl, quinazolinyl, 1,5-diazinyl, carbazole Base, benzodioxolyl, quinoxalinyl, purinyl, furanyl, isobenzylfuranyl (isobenzylfuranyl), benzimidazolyl, benzofuryl, benzothienyl, quinolyl Linyl, indolyl, isoquinolyl, dibenzofuryl, imidazo[1,2-α]pyridyl, [1,2,4-triazolo][4,3-α]pyridyl , pyrazolo[1,5-α]pyridyl, [1,2,4-triazolo][1,5-α]pyridyl, 2-oxo-1,3-benzoxazolyl, 4-oxo-3H-quinazolinyl, 3-oxo-[1,2,4]-triazolo[4,3-α]-2H-pyridyl, 5-oxo-[1,2 , 4]-4H-oxadiazolyl, 2-oxo-[1,3,4]-3H-oxadiazolyl, 2-oxo-1,3-dihydro-2H-imidazolyl, 3- Oxo-2,4-dihydro-3H-1,2,4-triazolyl, etc. For heterocyclyl and heteroaryl, rings and ring systems containing 3-15 atoms are included, forming 1-3 rings.

"Halogen" refers to fluorine, chlorine, bromine and iodine. Chlorine and fluorine are generally preferred. Fluorine (eg _CF3O and _CF3CH2O ) is most preferred when halogen is substituted on the alkyl or _alkoxy group.

The compounds of the present invention may contain one or more asymmetric centers and thus exist as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and single diastereomers. The compounds of the invention in formula Ia have an asymmetric center at the carbon atom marked with ^* . Depending on the nature of the various substituents on the molecule, additional centers of asymmetry may exist. Each such asymmetric center will independently give rise to two optical isomers, and all possible optical isomers and diastereomers, in mixtures and as pure or partially pure compounds, are included within the scope of the present invention. within range. The present invention includes all such isomeric forms of these compounds.

Certain compounds described herein contain olefinic double bonds and, unless otherwise indicated, are meant to include both E and Z geometric isomers.

Certain compounds described herein may exist as tautomers, which have different points of attachment of hydrogens with one or more double bond inversions. For example, a ketone and its enol form are keto-enol tautomers. Individual tautomers and mixtures thereof are included in the compounds of the present invention.

The classes of compounds shown in Formula I have no preferred stereochemistry. Formula Ia shows the preferred stereochemistry at the carbon atom attached to the amino group of the beta amino acid from which these compounds are prepared.

These diastereomers may be synthesized individually or separated chromatographically as known in the art by appropriate modification of the methodology disclosed herein. The absolute stereochemistry of the crystalline products or crystalline intermediates may be determined by X-ray crystallography or, if necessary, derivatized with reagents containing asymmetric centers of known absolute configuration.

Racemic mixtures of compounds can be separated, if desired, to isolate the individual enantiomers. Separation can be accomplished by methods well known in the art, such as coupling a racemic mixture of a compound to an enantiomerically pure compound to form a diastereomeric mixture, followed by separation of the individual diastereomeric compounds by standard methods such as fractional crystallization or chromatography. body. Coupling reactions are usually salt formation with enantiomerically pure acids or bases. Diastereomeric derivatives can then be converted to the pure enantiomers by cleavage of the added chiral residue. Racemic mixtures of compounds can be isolated directly or by chromatographic methods well known in the art using chiral stationary phases.

Alternatively, any enantiomer of a compound may be obtained by stereoselective synthesis using optically pure starting materials or reagents of known configuration by methods well known in the art.

It should be understood that, as used herein, compounds of structural formula I also include pharmaceutically acceptable salts, and non-pharmaceutically acceptable salts when used as precursors of the free compounds or their pharmaceutically acceptable salts or in other synthetic procedures. Accepted salt.

The compounds of the present invention may be administered in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids, including inorganic or organic bases and inorganic or organic acids. Salts of basic compounds encompassed within the term "pharmaceutically acceptable salt" refer to non-toxic salts of compounds of the present invention, usually prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts of basic compounds of the invention include, but are not limited to, the following salts: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, camphor Sulfonate (camsylate), carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, ettoate, ethanesulfonate, fumarate salt, glucoheptonate, gluconate, glutamate, glycolyl p-aminophenylarsinate, hexyl-2,4-dihydroxybenzoate (hexylresorcinate), hydrabamine (hydrabamine) ), hydrobromide, hydrochloride, xinanate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, almond salt, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, naphthalenesulfonate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate salt, pamoate (emboate), palmitate, pantothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, sulfate, Subacetate, succinate, tannate, tartrate, theanate, tosylate, triethiodide, and valerate. Additionally, when the compound of the present invention has an acidic moiety, suitable pharmaceutically acceptable salts thereof include, but are not limited to, salts derived from inorganic bases including: aluminum, ammonium, calcium, copper, ferric, ferrous , lithium, magnesium, trivalent manganese, divalent manganese, potassium, sodium, zinc, etc. Particular preference is given to the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include those of primary, secondary and tertiary amines; cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N, N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucose Glucosamine, glucosamine, histidine, hepamine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine , Theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

Furthermore, where a carboxylic acid (-COOH) or alcohol group is present in the compounds of the present invention, pharmaceutically acceptable esters of carboxylic acid derivatives such as methyl, ethyl or pivaloyloxymethyl esters may be used, or Acyl derivatives of alcohols such as acetates or maleates. Also included are esters and acyl groups known in the art to modify solubility or hydrolysis properties for use as sustained release or prodrug formulations.

Solvates, especially hydrates, of the compounds of formula I are also included in the present invention.

The invention is illustrated in the Examples and the use of the compounds disclosed herein.

Compounds of interest are useful in a method of inhibiting dipeptidyl peptidase-IV enzyme in a patient (eg, a mammal) in need of such inhibition comprising administering an effective amount of the compound. The present invention relates to the use of the compounds disclosed herein as inhibitors of dipeptidyl peptidase-IV enzyme activity.

In addition to primates such as humans, the methods of the invention can treat a variety of other mammals. For example, treatable mammals include, but are not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats, or other bovine, ovine, equine, canine, feline, rodent, or murine species. However, the methods of the invention can also be practiced in other species such as birds (eg chickens).

The present invention also relates to a method for the preparation of a medicament for inhibiting dipeptidyl peptidase-IV enzyme activity in humans and animals, the method comprising combining the compound of the present invention with a pharmaceutically acceptable carrier or diluent.

The subject to be treated by the methods of the invention is typically a mammal, preferably a human, male or female, in which inhibition of dipeptidyl peptidase-IV enzyme activity is desired. The term "therapeutically effective amount" refers to the amount of a compound of interest that produces a biological or medical response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, physician or other clinician.

As used herein, the term "composition" includes products comprising the specified ingredients in the specified amounts, as well as any product that results directly or indirectly from the combination of the specified ingredients in the specified amounts. This term in relation to pharmaceutical compositions includes products comprising the active ingredient and inert ingredients constituting the carrier, as well as any combination, complexation, or aggregation of two or more ingredients, or dissociation of one or more ingredients. , or any product resulting, directly or indirectly, from any other type of reaction or interaction of one or more ingredients. Accordingly, the pharmaceutical compositions of the present invention include any composition prepared by admixing a compound of the present invention and a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The term "administering" a compound should be understood as providing a compound of the invention or a prodrug of a compound of the invention to a subject in need of treatment.

The use of the compounds of the invention as inhibitors of dipeptidyl peptidase-IV enzyme activity can be demonstrated by methodologies known in the art. Inhibition constants were determined as follows. Sequential fluorescence assays are performed using the substrate Gly-Pro-AMC, which is cleaved by DP-IV to release the fluorescent AMC leaving group. The kinetic parameters describing the reaction are as follows: K _m =50 μM; k _cat =75 s ⁻¹ ; k _cat /K _m =1.5×10 ⁶ M ⁻¹ s ⁻¹ . A typical reaction contains approximately 50 pM enzyme, 50 μM Gly-Pro-AMC and buffer (100 mM HEPES, pH 7.5, 0.1 mg/ml BAS) in a total reaction volume of 100 μl. The release of AMC was continuously monitored on a 96-well plate fluorometer using an excitation wavelength of 360 nm and an emission wavelength of 460 nm. In this case, approximately 0.8 μM AMC was generated in 30 minutes at 25°C. The enzyme used in these studies was a soluble (except transmembrane domain and cytoplasmic extension) human protein produced in a baculovirus expression system (Bac-To-Bac, Gibco BRL). The kinetic constants for Gly-Pro-AMC and GLP-1 hydrolysis were found to be consistent with literature values for the native enzymes. To measure dissociation constants of compounds, inhibitors in DMSO were added to reactions containing enzyme and substrate (final concentration of DMSO was 1%). All experiments were performed at room temperature using the standard reaction conditions described above. To determine dissociation constants (K _i ), reaction rates were fitted by nonlinear regression to the Michaelis-Menton equation for competitive inhibition. Errors in reproducing dissociation constants are generally less than a factor of 2.

Specifically, the compounds of the following examples have activity in inhibiting dipeptidyl peptidase-IV enzymes in the above assays, with _IC50 generally less than about 1 μM. This result demonstrates the intrinsic activity of the compounds as inhibitors of dipeptidyl peptidase-IV enzyme activity.

Dipeptidyl peptidase-IV enzymes (DP-IV) are cell surface proteins involved in a wide range of physiological functions. It has a broad tissue distribution (gut, kidney, liver, pancreas, placenta, thymus, spleen, epithelial cells, vascular endothelium, lymphoid and myeloid cells, serum) and unique tissue and cell type expression levels. DP-IV is the same as T cell activation marker CD26, which can decompose various immunomodulatory peptides, endocrine peptides and neuropeptides in vitro. This suggests a potential role for this peptidase in various disease processes in humans or other species.

Accordingly, the subject compounds are useful in methods of preventing or treating the following diseases, disorders and conditions.

Type 2 diabetes and related disorders : It is well established that the incretins GLP-1 and GIP are rapidly inactivated by DP-IV in vivo. Studies and preclinical experiments with DP-IV ^(-l-) -deficient mice have shown that DP-IV inhibition increases steady-state concentrations of GLP-1 and GIP, leading to improved glucose tolerance. Similar to GLP-1 and GIP, it is possible that other glucagon family peptides involved in glucose regulation are also inactivated by DP-IV (eg PACAP). Inactivation of these peptides by DP-IV also plays a role in glucose homeostasis.

Accordingly, the DP-IV inhibitors of the present invention are useful in the treatment of type 2 diabetes, as well as in the treatment and prevention of a variety of conditions frequently associated with type 2 diabetes, including metabolic syndrome X, reactive hypoglycemia, and diabetic dyslipidemia. Obesity, discussed below, is another condition often found with type 2 diabetes that is responsive to treatment with the compounds of the invention.

The following diseases, disorders and conditions are associated with type 2 diabetes and thus can be treated, controlled or in some cases prevented by treatment with the compounds of the invention: (1) hyperglycemia, (2) low glucose tolerance, (3) Insulin resistance, (4) obesity, (5) lipid disorder, (6) dyslipidemia, (7) hyperlipidemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, ( 10) Low HDL level, (11) High LDL level, (12) Atherosclerosis and its sequelae, (13) Vascular restenosis, (14) Stress bowel syndrome, (15) Inflammatory bowel disease, Including Crohn's disease and ulcerative colitis, (16) other inflammatory diseases, (17) pancreatitis, (18) abdominal obesity, (19) neurodegenerative diseases, (20) retinopathy, (21) nephropathy, ( 22) Neuropathy, (23) Hypertension, (24) Syndrome X, (25) Ovarian hyperandrogenism (polycystic ovary syndrome) and other disorders of which insulin resistance is a component.

Obesity : DP-IV inhibitors are useful in the treatment of obesity. This is based on the observed inhibitory effects of GLP-1 and GLP-2 on food intake and gastric emptying. Administration of exogenous GLP-1 significantly reduces human food intake and slows gastric emptying ( Am. J. Physiol ., 277: R910-R916 (1999)). ICV administration of GLP-1 to rats and mice also had a significant effect on food intake ( Nature Medicine , 2: 1254-1258 (1996)). This inhibitory effect of feeding was not observed in GLP-1R(-l-) mice, suggesting that this effect is mediated by brain GLP-1 receptors. Similar to GLP-1, it is possible that GLP-2 is also regulated by DP-IV. ICV administration of GLP-2 also inhibits food intake, similar to the effect observed with GLP-1 ( Nature Medicine , 6:802-807 (2000)). In addition, studies with DP-IV deficient mice suggest that these animals are resistant to diet-induced obesity and related pathologies such as hyperinsulinonemia.

Growth hormone deficiency : Based on the hypothesis that growth hormone releasing factor (GRF), a peptide that stimulates growth hormone release from the anterior pituitary gland, is cleaved in vivo by the enzyme DP-IV (WO00/56297), DP-IV inhibition could be used in Treatment of growth hormone deficiency. The following data provide evidence that GRF is an endogenous substrate: (1) GRF is efficiently cleaved in vitro to generate inactivated product GRF[3-44] ( BBA 1122:147-153(1992)); Rapid degradation to GRF[3-44] in plasma; this was prevented by the DP-IV inhibitor diprotin A; (3) GRF[3-44] was found in the plasma of human GRF transgenic pigs ( J.Clin.Invest ., 83:1533-1540 (1989)). DP-IV inhibitors are therefore useful in the same spectrum of indications that have been considered indications for growth hormone secretagogues.

Intestinal Injury : Findings suggest the potential of using DP-IV inhibitors to treat intestinal injury, suggesting that glucagon-like peptide-2 (GLP-2), a possible endogenous substrate of DP-IV, may act on the intestinal epithelium. Presents nutritional effects ( Regulatory Peptides , 90:27-32 (2000)). Administration of GLP-2 resulted in increased small bowel mass in rodents and reduced intestinal damage in rodent models of colitis and enteritis.

Immunosuppression : Based on studies demonstrating the role of DP-IV enzymes in T cell activation and chemokine processing and the efficacy of DP-IV inhibitors in in vivo disease models, DP-IV inhibition may be used to modulate immune responses. DP-IV has been shown to be identical to CD26, a cell surface marker of activated immune cells. CD26 expression is regulated by the differentiation and activation status of immune cells. It is generally accepted that CD26 functions as a co-stimulatory molecule in in vitro models of T cell activation. Several chemokines contain a proline at the penultimate position, presumably to protect them from degradation by nonspecific aminopeptidases. Many of these chemokines have been shown to be processed by DP-IV in vitro. In some cases (RANTES, LD78-β, MDC, eotaxin, SDF-1α), cleavage resulted in altered activity in chemotaxis and signaling assays. In some cases (RANTES), receptor selectivity also appeared to be altered. A number of N-terminal truncated forms of various chemokines have been identified in in vitro cell culture systems, including predicted DP-IV hydrolysates.

DP-IV inhibitors have been shown to be effective immunosuppressants in transplantation and animal models of arthritis. Prodipine (Pro-Pro-diphenyl-phosphonate), an irreversible inhibitor of DP-IV, showed a doubling of cardiac allograft survival on days 7-14 in rats ( Transplantation , 63 : 1495-1500 (1997)). DP-IV inhibitors have been tested in collagen- and alkyldiamine-induced arthritis in rats and showed a statistically significant reduction in hindpaw swelling in this model [ Int.J, Immunopharmacology, 19:15-24( 1997) and Immunopharmacology, 40:21-26 (1998)]. DP-IV is upregulated in a variety of autoimmune diseases, including rheumatoid arthritis, multiple sclerosis, Graves' disease, and Hashimoto's thyroiditis ( Immunology Today , 20:367-375 (1999)).

HIV infection : DP-IV inhibition can be used to treat or prevent HIV infection or AIDS, because various chemokines that inhibit HIV cell entry are potential substrates of DP-IV ( Immunology Today , 20: 367-375 (1999)) . For SDF-la, cleavage reduces antiviral activity ( PNAS , 95:6331-6 (1998)). Therefore, stabilization of SDF-la by inhibiting DP-IV is expected to reduce HIV infectivity.

Hematopoiesis : Inhibition of DP-IV is useful in the treatment or prevention of disorders of hematopoiesis, as DP-IV can be associated with hematopoiesis. A DP-IV inhibitor, Val-Boro-Pro, stimulates hematopoiesis in a mouse model of cyclophosphamide-induced neutropenia.

Neuronal Disorders : DP-IV inhibition is useful in the treatment or prevention of a variety of neuronal or neurological disorders, as various peptides involved in various neuronal processes are cleaved by DP-IV in vitro. Accordingly, DP-IV inhibitors may have therapeutic benefit in the treatment of neuronal disorders. Endorphin-2, β-casomorphin and substance P have all been shown to be substrates of DP-IV in vitro. In all cases, in vitro lysis was efficient with k _cat /K _m of about 106 M ⁻¹ s ⁻¹ or greater. In an electric shock experimental model of analgesia in rats, DP-IV inhibitors showed significant effects independent of the presence of exogenous endorphin-2 ( Brain Research , 815:278-286 (1999)). The neuroprotective and neuroregenerative effects of DP-IV inhibitors were also evidenced by the ability of the inhibitors to: protect motor neurons from excitotoxic cell death, and protect striatal innervation of dopaminergic neurons when co-administered with MPTP , and promote restoration of striatal innervation density when given therapeutically following MPTP treatment [see Yong-Q. Wu et al., "Neuroprotective Effects of Inhibitors of Dipeptidyl Peptidase-IV InVitro," Int.Conf. Basic Science and Clinical Applications , 2002, September, 26-29 (Berlin, Germany)].

anxiety

Rats born deficient in DP-IV have an anxiolytic phenotype (WO 02/34243; Karl et al., Physiol. Behav . 2003). DP-IV-deficient mice also have an anxiolytic phenotype using the Porsolt swim test and the light/dark model. Accordingly, DP-IV inhibitors may prove useful in the treatment of anxiety and related disorders.

memory and cognition

GLP-1 agonists are active in models of learning (passive avoidance, Morris water maze) and neuronal injury (kainate-induced neuronal apoptosis) as demonstrated by During et al. ( Nature Med. 9:1173-1179 (2003)). This result suggests that GLP-1 has a physiological role in learning and neuroprotection. Stabilization of GLP-1 by DP-IV inhibitors is expected to show similar effects.

Tumor invasion and metastasis : DP-IV inhibition may be useful in the treatment or prevention of tumor invasion and metastasis, as increased or decreased expression of certain exopeptidases, including DP-IV, has been observed during transition of normal cells to a malignant phenotype ( J. Exp. Med., 190:301-305 (1999)). Upregulation or downregulation of these proteins appears to be tissue and cell type specific. For example, increased expression of CD26/DP-IV has been observed in T-cell lymphomas, T-cell acute lymphoblastic leukemia, cell-derived thyroid carcinomas, basal cell carcinomas, and breast cancers. Therefore, DP-IV inhibitors are useful in the treatment of this cancer.

Benign prostatic hypertrophy : DP-IV inhibition may be useful in the treatment of benign prostatic hypertrophy, as increased DP-IV activity was noted in prostate tissue from patients with BPH ( Eur.J. Clin.Chem.Clin.Biochem., 30: 333-338 (1992)).

Sperm motility/male contraception : DP-IV inhibition can be used to alter sperm motility and for male contraception because in semen the prostatosome (prostate-derived organelle important for sperm motility) has high levels of DP - IV activity ( Eur. J. Clin. Chem. Clin. Biochem., 30:333-338 (1992 )).

Gingivitis : DP-IV inhibition may be useful in the treatment of gingivitis, as DP-IV activity has been found in gingival crevicular fluid and has been associated with periodontal disease severity in some studies ( Arch. Oral Biol., 37:167 -173 (1992)).

Osteoporosis : DP-IV inhibition may be useful in the treatment or prevention of osteoporosis due to the presence of GIP receptors in osteoblasts.

The compounds of the present invention are useful in the treatment or prevention of one or more of the following conditions or diseases: (1) hyperglycemia, (2) low glucose tolerance, (3) insulin resistance, (4) obesity, (5) lipid disorders , (6) Dyslipidemia, (7) Hyperlipidemia, (8) Hypertriglyceridemia, (9) Hypercholesterolemia, (10) Low HDL level, (11) High LDL level, (12) Atherosclerosis and its sequelae, (13) Vascular restenosis, (14) Irritable bowel syndrome, (15) Inflammatory bowel disease, including Crohn's disease and ulcerative colitis, (16) Other Inflammatory Diseases, (17) Pancreatitis, (18) Abdominal Obesity, (19) Neurodegenerative Diseases, (20) Retinopathy, (21) Nephropathy, (22) Neuropathy, (23) Syndrome X, (24) Ovarian Hyperandrogenism (polycystic ovary syndrome), (25) type 2 diabetes, (26) growth hormone deficiency, (27) neutropenia, (28) neuronal disorders, (29) tumor metastasis , (30) benign prostatic hypertrophy, (32) gingivitis, (33) hypertension, (34) osteoporosis, and other conditions that can be treated or prevented by inhibiting DP-IV.

The compounds of interest are used in combination with other drugs, and are further used in methods of preventing or treating the above-mentioned diseases, disorders and conditions.

Compounds of the present invention may be used in combination with one or more other drugs for the treatment, prevention, suppression or alleviation of diseases or conditions for which compounds of formula I or other drugs may be useful, wherein the combination of said drugs is more effective than either drug alone. safe or effective. Said other drugs may be administered simultaneously or sequentially with the compound of formula I by their usual routes and amounts. When a compound of formula I is used concomitantly with one or more other drugs, a pharmaceutical composition comprising such other drugs and a compound of formula I in unit dosage form is preferred. However, combination therapy may also include therapies in which the compound of formula I and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the invention and other active ingredients may be used at lower dosages than when each is used alone. Accordingly, the pharmaceutical compositions of the present invention include those containing one or more other active ingredients in addition to the compound of formula I.

Other active ingredients administered in combination with the compound of formula I may be administered separately or in the same pharmaceutical composition, examples of which include but are not limited to:

(a) other dipeptidyl peptidase IV (DP-IV) inhibitors;

(b) Insulin sensitizers, including (i) PPARγ agonists such as glitazones (such as troglitazone, pioglitazone, emglitazone, MCC-555, rosiglitazone, etc.) and other PPAR ligands entities, including PPARα/γ dual agonists such as KRP-297, and PPARα agonists such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (ii) Biguanides such as metformin and phenformin, and (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors;

(c) insulin or insulin mimetics;

(d) sulfonylureas and other insulin secretagogues, such as tolbutamide, glibenclamide, glipizide, glimepiride, and meglitinides such as repaglinide;

(e) alpha-glucosidase inhibitors (such as acarbose and miglitol);

(f) glucagon receptor antagonists, such as disclosed in WO 98/04528, WO 99/01423, WO 00/39088 and WO 00/69810;

(g) GLP-1, GLP-1 mimetics and GLP-1 receptor agonists, such as disclosed in WO 00/42026 and WO 00/59887;

(h) GIP and GIP mimetics, such as disclosed in WO 00/58360, and GIP receptor agonists;

(i) PACAP, PACAP mimetics and PACAP receptor agonists, such as disclosed in WO01/23420;

(j) Cholesterol-lowering drugs such as (i) HMG-CoA reductase inhibitors (lovastatin, neovastatin, pravastatin, cerivastatin, fluvastatin, atorvastatin, itastatin, and rovastatin) Suvastatin and other statins), (ii) chelating agents (dialkylaminoalkyl derivatives of cholestyramine, colestipol, and cross-linked dextran), (iii) nicotinic alcohol, niacin or salts thereof, (iv) PPARα agonists such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (v) PPARα/γ dual agonists such as KRP-297, (vi) cholesterol absorption inhibitors such as β-sitosterol and ezetimibe, (vii) acyl-CoA:cholesterol acyltransferase inhibitors such as avasimibe, and (viii) antioxidants, e.g. Probucol;

(k) PPARδ agonists, such as disclosed in WO 97/28149;

(l) Anti-obesity compounds such as fenfluramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide _Y1 or _Y5 antagonists, CB1 receptor inverse agonists β3 _- adrenoceptor agonists, melanocortin receptor agonists especially melanocortin-4 receptor agonists, ghrelin antagonists and melanoconcentrating hormone (MCH) receptor Antagonist;

(m) ileal bile acid transport inhibitors;

(n) Drugs for inflammatory diseases, such as aspirin, nonsteroidal anti-inflammatory drugs, glucocorticoids, sulfasalazine, and selective cyclooxygenase-2 inhibitors;

(o) Antihypertensives such as ACE inhibitors (enalapril, lisinopril, captopril, quinapril, trandolapril), A-II receptor blockers (cloxapril, Tan, candesartan, irbesartan, valsartan, telmisartan, eprosartan), beta blockers and calcium channel blockers.

Dipeptidyl peptidase-IV inhibitors that can be combined with compounds of formula I include dipeptidyl peptidase-IV inhibitors disclosed in: WO 03/004498 (January 16, 2003), WO 03/004496 (2003 16 January), EP 1 258 476 (20 November 2002), WO 02/083128 (24 October 2002), WO 02/062764 (15 August 2002), WO 03/000250 ( 3 January 2003), WO 03/002530 (9 January 2003), WO 03/002531 (9 January 2003), WO 03/002553 (9 January 2003), WO 03/ 002593 (January 9, 2003), WO 03/000180 (January 3, 2003) and WO 03/000181 (January 3, 2003). Specific DP-IV inhibitor compounds include isoleucine thiazolidine, NVP-DPP728, P32/98 and LAF 237.

Anti-obesity compounds that can be combined with compounds of formula I include fenfluramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide _Y1 or _Y5 antagonists, cannabinoid CB1 Receptor antagonists or inverse agonists, melanocortin receptor agonists, especially melanocortin-4 receptor agonists, ghrelin antagonists and melanoconcentrating hormone (MCH) receptor antagonists. For a review of anti-obesity compounds that can be combined with compounds of formula I, see S. Chaki et al., "Recent advances in feeding suppressing agents: potential therapeutic strategy for the treatment of obesity," Expert Opin. Ther. Patents, 11: 1677- 1692 (2001) and D. Spanswick and K. Lee, "Emerging antiobesity drugs," Expert Opin. Emerging Drugs, 8: 217-237 (2003).

Neuropeptide Y5 antagonists that may be combined with compounds of formula I include those disclosed in U.S. Pat. The compounds are called GW 59884A, GW569180A, LY366377 and CGP-71683A.

Cannabinoid CB1 receptor antagonists that can be combined with the compound of formula I include those disclosed in PCT Publication WO 03/007887; U.S. Patent No. 5,624,941, such as rimonabant; PCT Publication WO 02/ 076949, such as SLV-319; US Patent No. 6028084; PCT Publication WO 98/41519; PCT Publication WO 00/10968; PCT Publication WO 99/02499;

Melanocortin receptor agonists that may be combined with compounds of formula I include those disclosed in WO 03/009847 (February 6, 2003), WO 02/068388 (September 6, 2001 ), WO 99/64002 (December 16, 1999), WO 00/74679 (December 14, 2000), WO 01/70708 (September 27, 2001), WO 01/70337 (September 2001 27) and JDSpeake et al., "Recent advances in the development of melanocortin-4 receptor agonists," Expert Opin. Ther. Patents , 12:1631-1638 (2002).

The above combinations include not only combinations of the compound of the present invention with one other active compound, but also combinations with two or more other active compounds. Non-limiting examples include combinations of compounds having formula I with two or more active compounds selected from the group consisting of biguanides, sulfonylureas, HMG-CoA reductase inhibitors, PPAR agonists, PTP-1B inhibitors, other DP-IV inhibitors and anti-obesity compounds.

Likewise, the compounds of the invention may be used in combination with other drugs for the treatment/prevention/suppression or amelioration of diseases or conditions for which the compounds of the invention are useful. Such other drugs may be administered simultaneously or sequentially with the compound of the present invention by the usual route and amount. When the compound of the present invention is used concomitantly with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include pharmaceutical compositions that also contain one or more other active ingredients in addition to a compound of the present invention.

The weight ratio of the compound of the present invention to the second active ingredient may vary depending on the effective dosage of each ingredient. An effective dose of each ingredient will generally be used. Thus, for example, when a compound of the present invention is combined with another drug, the weight ratio of the compound of the present invention to the other drug is generally in the range of about 1000:1 to 1:1000, preferably about 200:1 to 1:200 In the range. Combinations of the compounds of the present invention with other active ingredients are also generally within the above ranges, but in each case an effective dose of each active ingredient should be used.

In such combinations, the compound of the invention and the other active drug may be administered separately or together. Alternatively, one component may be administered prior to, simultaneously with, or subsequent to the administration of the other drug.

Compounds of the invention may be administered orally, parenterally (e.g., intramuscularly, intraperitoneally, intravenously, ICV, cisternal injection or bolus injection, subcutaneous injection or implant), by inhalation spray, nasal, vaginal, rectal, sublingual or topically They can be administered by various routes of administration, and can be formulated individually or together into unit dosage forms containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and excipients suitable for various routes of administration. In addition to the treatment of warm-blooded animals such as mice, rats, horses, cattle, sheep, dogs, cats, monkeys, etc., the compounds of the present invention are effective in humans.

The pharmaceutical compositions for administering the compounds of the present invention may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general, pharmaceutical compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired formulation. The compound of interest is included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the disease process or condition. As used herein, the term "composition" includes products comprising the specified ingredients in the specified amounts, as well as any product that results directly or indirectly from the combination of the specified ingredients in the specified amounts.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral use, for example as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs. The composition for oral administration can be prepared according to any method known in the field of preparation of pharmaceutical compositions, and this composition can contain one or more excipients selected from sweeteners, flavoring agents, coloring agents and preservatives, To provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents such as cornstarch or alginic acid; binders such as starch, gelatin or arabic acid; gums; and lubricants, such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated, or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thus provide sustained action over a longer period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. They may also be coated by the techniques described in US Pat. Nos. 4,256,108, 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

Formulations for oral administration may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily medium such as peanut oil, Mixed with liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and acacia; dispersing or wetting agents It may be a naturally occurring phospholipid, such as lecithin, or a condensation product of alkylene oxide with a fatty acid, such as polyoxyethylene stearate, or a condensation product of ethylene oxide with a long-chain fatty alcohol, such as heptadecylethoxyhexadecane Alkanol (heptadecaethyleneoxycetanol), or the condensation product of ethylene oxide with partial esters derived from fatty acids and hexitols such as polyoxyethylene sorbitan monooleate, or partial esters of ethylene oxide with fatty acids and hexitol anhydrides Condensation products of esters such as polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate; one or more coloring agents; one or more flavoring agents and one or more sweeteners. Flavoring agents such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening and flavoring agents such as those set forth above may be added to provide a palatable oral preparation. These compositions can be preserved by the addition of antioxidants such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified above. Additional adjuvants, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as liquid paraffin, or mixtures thereof. Suitable emulsifiers may be naturally occurring gums such as acacia or tragacanth; naturally occurring phospholipids such as soybean lecithin; esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate and condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsifiers may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents.

Pharmaceutical compositions can be in the form of sterile injectable aqueous or oily suspensions. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of this invention may also be administered in the form of suppositories for rectal administration. Such compositions are prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and therefore will melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, gels, solutions or suspensions, etc., containing the compounds of the invention may be employed. (For purposes of this use, topical use shall include mouthwashes and mouthwashes).

The pharmaceutical compositions and methods of the invention may further comprise other therapeutically active compounds as indicated herein which are commonly used in the treatment of the above-mentioned pathological conditions.

In the treatment or prevention of conditions requiring inhibition of dipeptidyl peptidase-IV enzyme activity, suitable dosage levels are generally about 0.01-500 mg per kg patient body weight per day, administered in single or multiple doses. Preferred dosage levels are about 0.1-250 mg/kg per day, more preferably about 0.5-100 mg/kg per day. Suitable dosage levels may be about 0.01-250 mg/kg per day, about 0.05-100 mg/kg per day, or about 0.1-50 mg/kg per day. Within this range the dosage may be 0.05-0.5, 0.5-5 or 5-50 mg/kg per day. For oral administration, the composition is preferably administered in the form of a tablet containing 1.0-1000 mg of active ingredient, especially 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 mg of active ingredient, adjusting the dosage for the treated patient according to the symptoms. The compound dosage regimen may be 1-4 times per day, preferably 1 or 2 times per day.

When treating or preventing diabetes and/or hyperglycemia or hypertriglyceridemia or other diseases for which the compound of the present invention is applicable, when the compound of the present invention is administered in a daily dose of about 0.1 mg-100 mg per kg of animal body weight, preferably as a single dose per day Satisfactory results are generally obtained when administered in single doses or in divided doses 2-6 times daily or in sustained release form. For most large mammals, the total daily dosage will be from about 1.0 mg to 1000 mg, preferably from about 1 mg to 50 mg. In the case of a 70 kg adult, the total daily dose will generally be about 7 mg to 350 mg. The dosage regimen can be adjusted to provide the optimum therapeutic response.

It is to be understood, however, that the specific dosage level and frequency of dosing for any particular patient may vary, depending on a variety of factors, including the activity of the particular compound employed, the metabolic stability and length of action of the compound, age, body weight , general health, sex, diet, mode and timing of administration, rate of excretion, drug combination, severity of specific condition and host being treated.

Certain methods for preparing compounds of the invention are illustrated in the following schemes and examples. Starting materials are prepared according to methods known in the art or as set forth herein.

Compounds of the invention can be prepared from alpha-amino acid intermediates of formula II and substituted heterocyclic intermediates of formula III using standard peptide coupling conditions followed by deprotection.

wherein m, p, W, X, Z, R ¹ , R ² , R ³ and R ⁴ are as defined above, and P is a suitable nitrogen protecting group such as tert-butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz ) or 9-fluorenylmethoxycarbonyl (Fmoc).

The preparation of these intermediates is described in the schemes below.

Process 1

(Ar = suitably substituted phenyl)

Compounds of formula II are commercially available, known in the literature or readily prepared by a variety of methods familiar to those skilled in the art. A convenient route described in the literature (X. Qian et al., Tetrahedron 1995, 51, 1033-1054) is described in Scheme 1. Treatment of an activated acid derivative such as an acid chloride 1 (which is commercially available or readily prepared from the corresponding acid by, for example, treatment with thionyl chloride or oxalyl chloride) with lithium phenyloxazolidinone 2 affords the acyl oxazolidinone oxazolidinone3 . Conjugate addition of oxazolidinone 3 to an appropriate aryl Grignard reagent 4 affords intermediate 5 . The α-azido moiety can be introduced in one of two convenient ways. First, boron enolate is generated from acyl oxazolidinone 5 by treatment with boron triflate and a base such as triethylamine or N,N-diisopropylethylamine, via reaction with N-succinimide , boron enolate is brominated. Substitution of the resulting bromide with an azide, for example by treatment with tetramethylguanidine azide, affords the azide 6 . Alternatively, the potassium enolate of the acyl oxazolidinone 5 , for example from potassium hexamethyldisilazide, can be reacted with 2,4,6-triisopropylbenzenesulfonyl azide to give the azide 6 directly. The azide is reduced by catalytic hydrogenation or by treatment with triphenylphosphine and the resulting amine is protected with a suitable group, for example by treatment with di-tert-butyl dicarbonate to its N-tert-butoxycarbonyl (Boc )derivative. Hydrolysis of the oxazolidinone is conveniently carried out by treatment with lithium hydroperoxide to afford the desired acid intermediate II. It is readily apparent to those skilled in the art that, via this route, acyl oxazolidinone 5 is converted to azide 6 by appropriate selection of the (R) or (S) enantiomer of oxazolidinone 2 and using the appropriate method, all four diastereomers of acid II can be obtained in enantiomerically pure form.

Process 2

Aryl and ^R2 substituents can be introduced in intermediate 5 in reverse order as illustrated in Scheme 2. Acid 8 is commercially available or readily prepared by a variety of methods known to those skilled in the art. In this approach, methyl acrylate ( 7 ) is treated with an appropriately substituted iodobenzene (ArI) under Stille coupling conditions to afford acid 8 after ester hydrolysis. The acid is activated, for example, by treatment with oxalyl chloride as its acid chloride or by reaction with pivaloyl chloride (PivCl) as its mixed anhydride, followed by treatment with lithium oxazolidinone 2 to give acyl oxazolidinone 9 . Copper-catalyzed addition with an appropriate Grignard reagent 10 affords the desired intermediate 5 . Transformation of intermediate II can be carried out as described in scheme 1.

Process 3

Another method for the preparation of intermediate II is shown in Scheme 3, wherein ^R2 contains an optionally substituted alkenyl, and ^R2 and the protected amine are mutually exclusive. Acid 8 can undergo EDC-mediated coupling with N,O-dimethylhydroxylamine followed by treatment with an appropriate Grignard reagent 11 to afford ketone 12 . Treatment with catecholboranes in an asymmetric fashion in the presence of the (R) isomer of, for example, CBS catalysts can be reduced to alcohols as described by EJCorey in Tetrahedron Lett. 36:9153-9156 (1995) 13 . Coupling of the alcohol with N-Boc glycine affords ester 14 . The [3,3]-σ shifted rearrangement of the enolate of ester 14 gives the intermediate as described by U. Kazmaier in Angew . IIa.

Process 4

Compounds of formula III are commercially available, known in the literature or readily prepared by a variety of methods familiar to those skilled in the art. A convenient method for the preparation of intermediate III is shown in Scheme 4, where X is CHF and W and Z are _CH2 . Suitably protected alcohols 15 are known in the literature or can be conveniently prepared by a variety of methods familiar to those skilled in the art by using fluorinating reagents such as (diethylamino)sulfur trifluoride (DAST) or trifluorinated Treatment with fluorinated [bis(2-methoxyethyl)amino]sulfur ( 16 ) and deprotection gives the fluorinated intermediate IIIa.

Process 5

A method for the preparation of intermediate III, wherein X is CF ₂ , and W and Z are CH ₂ , is shown in Scheme 5. The appropriately protected alcohol 16 is oxidized to the corresponding ketone 17 by various methods known to those skilled in the art. Treatment of ketone 17 with a fluorinating reagent such as DAST affords the fluorinated intermediate IIIb after deprotection.

Process 6

As shown in Scheme 6, under standard peptide coupling conditions, for example, using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 1-hydroxybenzotriazole (EDC/HOBT ) or hexafluorophosphate O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium and 1-hydroxy-7-azabenzotriazole (HATU/HOAT), intermediate II is coupled with III in a solvent such as N,N-dimethylformamide (DMF) or dichloromethane at ambient temperature for 3-48 hours to afford intermediate 18 . In some cases intermediate III may be a salt such as hydrochloride or trifluoroacetate, in which case it is convenient to add a base to the coupling reaction, usually N,N-diisopropylethyl amine. The protecting group is then removed, for example with trifluoroacetic acid or methanolic hydrogen chloride in the case of Boc, to give the desired amine I. If necessary, the product is purified from unwanted by-products by recrystallization, trituration, preparative thin layer chromatography, silica gel flash chromatography eg Biotage® apparatus or HPLC. Compounds purified by HPLC can be isolated as corresponding salts. The intermediate was purified in the same way.

In some cases, products I prepared as described in Scheme 6 can be further modified, for example by manipulating substituents on Ar or ^R2 . Such treatments include, but are not limited to, reduction, oxidation, alkylation, acylation and hydrolysis reactions, which are generally known to those skilled in the art.

Process 7

(Ar'=WGK suitable aryl, heteroaryl or heterocyclyl)

An example of this is illustrated in Scheme 7. Following the above-mentioned scheme for the synthesis of intermediate 18 , intermediate 19 can be obtained, wherein the substituent of phenyl is halogen, such as bromine and iodine. Coupling of bromide or iodide 19 with boronic acid 20 under Suzuki conditions in the presence of a palladium catalyst affords intermediate 18 . This was converted to product I as described in Scheme 6.

Process 8

Another such example is set forth in Scheme 8. Intermediate 19 was converted to the corresponding boronic acid ester 21 . Borate esters 21 can undergo Suzuki coupling with a suitable halide such as Ar'Br 22 in the presence of a palladium catalyst to afford biaryl derivatives 18 . This was converted to product I as described in Scheme 6.

Process 9

Scheme 9 illustrates an example where the ^R2 substituent undergoes further reaction. Ozonolysis of intermediate 18a followed by oxidation affords acid 18b . Acids can be coupled with amines to afford amides 18c . Intermediates 18b and 18c can be converted to products I as described in Scheme 6.

In some cases, the order in which the above reaction schemes are carried out can be varied to facilitate the reaction or to avoid undesired reaction products. The following examples are provided so that the invention may be more fully understood. These examples are illustrative only and should not be construed as limiting the invention in any way.

Intermediate 1

(3S)-3-fluoropyrrolidine hydrochloride

Step A: Benzyl (3R)-3-hydroxypyrrolidine-1-carboxylate

To a 22-L 3-neck round bottom flask equipped with a mechanical stirrer, thermocouple, addition funnel, and nitrogen diffuser was added 425 g (4.88 mol) of (3R)-3-hydroxypyrrolidine, 8 L of dichloromethane, and 1 L of (7.17 mol) triethylamine. The solution was cooled to 5-10°C with an ice bath, and then 1000 g (5.86 mol) of benzyl chloroformate was added dropwise within about 1.5 hours, keeping the reaction temperature <20°C. The reaction mixture was stirred for an additional 1 hour in the ice bath, then the ice bath was removed and the reaction mixture was allowed to warm to ambient temperature overnight. The mixture was poured into a large extractor containing about ~15 L of saturated aqueous sodium bicarbonate. The aqueous phase was back extracted with 2 portions of 2-L dichloromethane. The combined organics were dried over magnesium sulfate and concentrated to an orange oil. The crude product was treated with dichloromethane, applied to a 5-kg silica gel column prepacked with 50% ethyl acetate/hexane, and eluted sequentially with 8 L of 50%, 16 L of 75%, and 100% ethyl acetate/hexane to give The title compound is a yellow oil which crystallizes on standing.

Step B: Benzyl (3S)-3-fluoropyrrolidine-1-carboxylate

To a 5-L 3-neck round bottom flask equipped with a mechanical stirrer, thermocouple, addition funnel, and nitrogen diffuser was added 375 mL (2.84 mol) of (diethylamino)sulfur trifluoride and 400 mL of dichloromethane. The solution was cooled to -78°C. A solution of 304 g (1.37 mol) (3R)-benzyl 3-hydroxypyrrolidine-1-carboxylate in 400 mL of dichloromethane was added via an addition funnel within 2 hours, keeping the reaction temperature <-70°C. The reaction mixture was stirred and allowed to warm slowly to ambient temperature overnight. The reaction mixture was added carefully in portions to a large extractor containing ice, water and saturated aqueous sodium bicarbonate. The mixture was extracted with 8 L of ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated to give a brown oil. Purification by flash chromatography (silica gel, gradient elution with 10-30% ethyl acetate/hexanes) afforded the title compound as a brown oil.

Step C: (3S)-3-fluoropyrrolidine hydrochloride

Benzyl (3S)-3-fluoropyrrolidine-1-carboxylate (249 g, 1.11 mmol) was dissolved in 2.3 L of ethanol, then 115 mL of water was added, followed by 30 g of 10% palladium on carbon. The mixture was shaken under 40 psi hydrogen for about 24 hours. An additional 10 g and then 5 g of catalyst were added successively. The mixture was stirred under 40 psi of hydrogen until the reaction was complete. The mixture was filtered and the filter cake was washed with ethanol. The filtrate and washings were combined, treated with 185 mL of concentrated hydrochloric acid, and concentrated to a colorless oil. The residue was azeotroped with toluene, then 2L of diethyl ether was added. Add isopropanol until the oil crystallizes. Aging the mixture at room temperature over the weekend. The crystals were collected, washed with ether, and dried in vacuo to give the title compound. [α] _D = +8.64 (c = 4, methanol).

Intermediate 2

(3R)-3-fluoropyrrolidine hydrochloride

Step A: Benzyl (3S)-3-acetoxypyrrolidine-1-carboxylate

To a 22-L 3-neck round bottom flask equipped with a mechanical stirrer, thermocouple, addition funnel, and nitrogen diffuser was added 422 g (1.91 mol) of benzyl (3R)-3-hydroxypyrrolidine-1-carboxylate (middle Body 1, step A), 12 L of toluene, 751 g (2.86 mol) of triphenylphosphine and 164 mL (2.86 mol) of glacial acetic acid. The resulting mixture was stirred at ambient temperature, then 500 g (2.87 mol) of diethyl azodicarboxylate was added via the addition funnel over about 30 minutes, maintaining the internal temperature <28°C with a cold water bath. The reaction was stirred overnight at ambient temperature. The solvent was removed in vacuo and the residue was triturated with 6 L of ether. The solid was filtered off and washed carefully with ether. The combined filtrate and ether washings were concentrated to a viscous yellow oil with solids. Purification by flash chromatography (silica gel, eluting with a gradient of 5% and then 10% to 30% ethyl acetate/hexanes) afforded the title compound as a pale yellow oil.

Step B: Benzyl (3S)-3-hydroxypyrrolidine-1-carboxylate

To the 20-L 3-neck round bottom flask containing 427g (1.62mol) (3S)-3-acetoxypyrrolidine-1-carboxylic acid benzyl ester, successively add 4L absolute ethanol and 101g (1.57mol) potassium hydroxide 400mL aqueous solution. After about 15 minutes, the reaction mixture was poured into 8 L of water and extracted with 8 L of ethyl acetate. The aqueous layer was then extracted with another 4 L of ethyl acetate. The combined organics were washed with saturated brine, dried over magnesium sulfate, and concentrated to a thick oil and solid.

Step C: Benzyl (3R)-3-fluoropyrrolidine-1-carboxylate

Following essentially the procedure outlined in Intermediate 1, Step B, 366 g (1.62 mol) of a portion of benzyl (3S)-3-hydroxypyrrolidine-1-carboxylate were converted to the title compound.

Step D: (3R)-3-Fluoropyrrolidine hydrochloride

Following essentially the procedure outlined in Intermediate 1, Step C, 222 g (1.0 mol) of a portion of benzyl (3R)-3-fluoropyrrolidine-1-carboxylate was converted to the title compound. [α] _D = -8.61 (c = 4, methanol).

Intermediate 3

3,3-Difluoropyrrolidine hydrochloride

Step A: Benzyl 3-oxopyrrolidine-1-carboxylate

To a 12-L 3-necked round bottom flask equipped with a mechanical stirrer, thermocouple, condenser, and nitrogen diffuser was added 351 g (1.61 mol) of benzyl (3R)-3-hydroxypyrrolidine-1-carboxylate (middle Body 1, step A), 6 L of dichloromethane, 500 g of powdered molecular sieves and 400 g (3.41 mol) of N-methylmorpholine-N-oxide. The resulting suspension was stirred at ambient temperature, to which 12.9 g (0.0367 mol) of tetrapropylammonium perruthenate were added. Keep the reaction temperature at ≤30°C with a cold water bath. The mixture was stirred at ambient temperature for 2 hours. The mixture was placed on a 5 kg silica gel column and eluted with 10% ethyl acetate/dichloromethane to give the title compound as an orange oil.

Step B: Benzyl 3,3-difluoropyrrolidine-1-carboxylate

To a 12 L 3 neck round bottom flask equipped with a mechanical stirrer, thermocouple, addition funnel and nitrogen diffuser was added 292 g (1.33 mol) of benzyl 3-oxopyrrolidine-1-carboxylate and 3 L of dichloromethane. At ambient temperature, 530 mL (4.0 mol) of (diethylamino)sulfur trifluoride was added dropwise to the stirred solution over 3 hours, maintaining the internal temperature below 25°C with a cold water bath. The mixture was stirred overnight at ambient temperature. The mixture was poured into a large extractor containing ice and solid sodium bicarbonate. Then 8 L of ethyl acetate was added and the mixture was basified with sodium bicarbonate. The organic layer was dried over magnesium sulfate and concentrated to 309 g of a brown oil. Purification by flash chromatography (silica gel, 10-20% ethyl acetate/hexanes gradient) afforded the title compound.

Step C: 3,3-difluoropyrrolidine hydrochloride

Following essentially the procedure outlined in Intermediate 1, Step C, 242 g (1.00 mol) of a portion of benzyl 3,3-difluoropyrrolidine-1-carboxylate were converted to the title compound. ¹ H NMR (500 MHz, CD ₃ OD): δ 3.7 (t, 2H), 3.6 (t, 2H), 2.55 (m, 2H).

Intermediate 4

4-fluoropiperidine hydrochloride

Step A: Benzyl 4-fluoro-1-piperidinecarboxylate

To a 1-L round bottom flask was added 12.64 g (51.4 mmol) benzyl 4-oxo-1-piperidinecarboxylate and 300 mL dichloromethane. To the stirred solution at -78°C was added 19 mL (102.8 mmol) of [bis(2-methoxyethyl)amino]sulfur trifluoride via an addition funnel over about 1 hour. The reaction mixture was allowed to warm slowly to ambient temperature overnight. The reaction mixture was added carefully in portions to a large extractor containing water and saturated aqueous sodium bicarbonate. The mixture was extracted with dichloromethane (3 x 300 mL). The combined organic layers were washed once with saturated aqueous sodium bicarbonate, twice with 10% aqueous hydrochloric acid and saturated brine, dried over sodium sulfate, and concentrated in vacuo. Purification by flash chromatography (gradient, hexane-65% ethyl acetate/hexane) on a Biotage® system gave the desired product. LC/MS 242.1 (M+1).

Step B: 4-fluoropiperidine hydrochloride

Benzyl 4-fluoro-1-piperidinecarboxylate (5.5 g, 23.2 mmol) was dissolved in 80 mL of ethanol, and 1.0 g of 20% palladium hydroxide (dry basis) on charcoal was added to the mixture. The mixture was shaken under 40 psi of hydrogen for about 12 hours, then filtered through a pad of celite, washing with 100 mL of methanol. The filtrate and washings were combined, treated with 60 mL of 1M hydrochloric acid in ether, and concentrated to a white waxy solid. The solid was dried in vacuo to give the title compound as a solid. The material was used without further purification. ¹ H NMR (CDCl ₃ ): δ4.95 (d, J=47.4Hz, 1H), 3.70 (br s, 1H), 3.34-3.27 (m, 4H), 2.29 (dt, J=37.1, 12.3Hz, 2H), 2.16 (br s, 2H).

Intermediate 5

3-Fluoroazetidine trifluoroacetate

Step A: 1-Benzhydryl-3-fluoroazetidine

To a 250 mL round bottom flask was added 3.0 g (12.5 mmol) of 1-benzhydryl-3-fluoroazetidine and 80 mL of dichloromethane. To the stirred solution at -78°C was added 4.6 mL (25 mmol) of [bis(2-methoxyethyl)amino]sulfur trifluoride via an addition funnel over about 3 hours. The reaction mixture was allowed to warm slowly to ambient temperature overnight. The reaction mixture was added portionwise (cautiously) to a large extractor containing water and saturated aqueous sodium bicarbonate. The mixture was extracted three times with 80 mL of dichloromethane. The combined organic layers were washed successively with saturated aqueous sodium bicarbonate solution, water and saturated brine, dried over sodium sulfate, and concentrated in vacuo. Purification using Biotage® system flash chromatography (gradient, hexanes - 80% ethyl acetate/hexanes) afforded the desired product. LC/MS 242.1 (M+1).

Step B: 3-Fluoroazetidine trifluoroacetate

1-Benzhydryl-3-fluoroazetidine (1.7 g, 7.04 mmol) was dissolved in 60 mL of ethanol and 500 mg of 20% palladium hydroxide (dry basis) on charcoal was added. The mixture was shaken under 40 psi of hydrogen for about 12 hours. The mixture was filtered through a pad of celite, and the filter cake was washed with 100 mL of methanol. The combined washes were treated with 10 mL of trifluoroacetic acid and concentrated to give two oils, the thicker being the desired trifluoroacetic acid salt. The mixture was used without further purification. ¹ H NMR (CDCl ₃ ) δ 5.45-4.30 (dm, J=56.7Hz, 1H), 4.46-4.38 (m, 2H), 4.24-2.17 (m, 2H).

Example 1

(3S)-1-[2S,3S)-2-amino-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1-oxobutyl]-3-fluoropyrrole Alkane trifluoroacetate

Step A: (4R)-3-[(2E)-3-(4-Bromophenyl)prop-2-enoyl]-4-phenyl-1,3-oxazolidin-2-one

To a stirred solution of 4-bromocinnamic acid (5.79 g, 22.5 mmol) in anhydrous THF (250 mL) was added successively triethylamine (4.60 mL, 34.6 mmol) and trimethylacetyl chloride ( 3.54 mL, 24.7 mmol). The resulting suspension was stirred at -78 °C for 15 min, at 0 °C for 1 h, then at -78 °C for 15 min before cannula transfer to 4(R)-4-phenyl at 0 °C - In the slurry of lithium 2-oxazolidinone, the latter was added to 4(R)-4-phenyl-2-oxazolidinone (5.0g, 30.6mmol) at -78°C 15 minutes ago Prepare by adding n-butyllithium (19.1 mL, 30.5 mmol) to anhydrous THF (150 mL) solution. The resulting slurry was stirred at -78°C for 1 hour and at room temperature for 12 hours. The reaction was quenched with saturated aqueous ammonium chloride. The organic phase was separated, concentrated in vacuo, and the crude product was used directly in the next step. LC/MS 370.2 (M+1).

Step B: (4R)-3-[(3R)-3-(4-Bromophenyl)butyryl]-4-phenyl-1,3-oxazolidin-2-one

To a stirred solution of copper(II) bromide dimethylsulfide complex (8.78 g, 42.7 mmol) in THF (60 mL) and dimethylsulfide (30 mL) was added methylmagnesium bromide ( 12.7 mL, 3.0 M in ether, 38.1 mmol). The resulting mixture was stirred at -40°C for 30 minutes and then allowed to warm to -20°C. The product of Step A (3.53 g, 9.48 mmol) in THF (30 mL) was added to the above reaction mixture at -20°C over 1 hour. The resulting mixture was stirred at -20°C for 2 hours, then slowly warmed to room temperature and stirred at room temperature for 12 hours. The reaction was quenched by the slow addition of saturated aqueous ammonium chloride. The organic phase was separated and the aqueous phase was extracted with two portions of ethyl acetate. The combined organic layers were washed with saturated brine and concentrated in vacuo. Purification by flash chromatography (silica gel, 83:17 hexane/ethyl acetate) afforded the desired product.

Step C: (4R)-3-[(2R,3S)-2-bromo-3-(4-bromophenyl)butyryl]-4-phenyl-1,3-oxazolidine- 2-keto

To a stirred solution of the product from Step B (2.87 g, 7.39 mmol) in dichloromethane (40 mL) was added diisopropylethylamine (1.93 mL, 11.1 mmol) and trifluoromethanesulfonic acid at -78°C Dibutylboron (9.6 mL, 1M in dichloromethane, 9.6 mmol). The light yellow solution was stirred at -78°C for 15 minutes, at 0°C for 1 hour, cooled again to -78°C and stirred for 15 minutes. The above solution was transferred via cannula to a pre-cooled suspension of N-bromosuccinimide (3.93 g, 22.2 mmol) in dichloromethane (40 mL). The resulting mixture was stirred at -78°C for 1 hour and at 0°C for 3 hours. The reaction was quenched by the addition of 0.5N aqueous sodium bisulfite. The organic phase was separated and the aqueous phase was extracted with two portions of ethyl acetate. The combined organic layers were washed with saturated brine and concentrated in vacuo. Purification by flash chromatography (silica gel, 83:17 hexane/ethyl acetate) afforded the desired product.

Step D: (4R)-3-[(2S,3S)-2-Azido-3-(4-bromophenyl)butyryl]-4-phenyl-1,3-oxa oxazolidin-2-one

To a stirred solution of the product from Step C (2.71 g, 6.39 mmol) in acetonitrile (40 mL) was added tetramethylguanidine azide (3.51 g, 22.2 mmol). The reaction was stirred at room temperature for 12 hours. The solid was filtered off and the filtrate was evaporated. The crude product was purified by flash chromatography (83:17 hexane/ethyl acetate) to give the desired product.

Step E: (2S,3S)-2-Azido-3-(4-bromophenyl)butanoic acid

To a stirred solution of the product from Step D (2.77 g, 6.23 mmol) in THF (60 mL) was added water (20 mL). The solution was stirred at 0° C. for 15 minutes, then 30% hydrogen peroxide (6.0 mL, 52.9 mmol) was added followed by lithium hydroxide (0.50 g, 21.2 mmol) slowly. The resulting mixture was stirred at 0°C for 4 hours. The reaction was quenched by the addition of saturated aqueous sodium sulfite and stirred at room temperature for 30 minutes. The aqueous phase was separated and washed with three portions of dichloromethane. The aqueous phase was then acidified to pH 1 with 3N hydrochloric acid and extracted with three portions of ethyl acetate. The ethyl acetate extracts were combined, dried over sodium sulfate and evaporated in vacuo to give the product which was used directly in the next step.

Step F: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-(4-bromophenyl)-1-oxo Butyl]-3-fluoropyrrolidine

To 1.20 g (4.22 mmol) of acid dissolved in anhydrous DMF (10 mL) was added EDC (2.29 g, 11.9 mmol), HOBT (1.62 g, 11.9 mmol), (3S)-3-fluoropyrrolidine hydrochloride (1.50 g, 11.9 mmol) and N,N'-diisopropylethylamine (4.2 mL, 23.6 mmol). After stirring at room temperature for 12 hours, the reaction was diluted with ethyl acetate. The organic phase was washed successively with saturated brine, 1N hydrochloric acid and 1N aqueous sodium hydroxide solution, dried over sodium sulfate and evaporated in vacuo to give a yellow foam. To this foam was added 40 mL of dioxane, 4 mL of water and triphenylphosphine (4.70 g, 17.9 mmol). The reaction was heated at 90°C for 12 hours and then cooled to room temperature. The solvent was removed in vacuo and the residue was dissolved in 20 mL of dioxane and 20 mL of saturated aqueous sodium bicarbonate. To the resulting mixture was added 7.8 g of di-tert-butyl dicarbonate (35.8 mmol). The reaction was stirred at room temperature for 12 hours. The reaction mixture was diluted with ethyl acetate and acidified to pH 1 with 1N hydrochloric acid. The layers were separated and the aqueous layer was extracted with two portions of ethyl acetate. The organic extracts were combined, washed with saturated brine, dried over sodium sulfate, and concentrated in vacuo. Purification by flash chromatography (silica gel, 66:34 hexane/ethyl acetate) afforded the desired product. LC/MS 429.1 (M+1).

Step G: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-(4'-fluoro-1,1'-biphenyl-4- Base)-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Step F (51.0 mg, 0.12 mmol) in 2 mL of toluene and 2 mL of ethanol was added 4-fluorophenylboronic acid (49.0 mg, 0.29 mmol), 1,1'-bis(diphenylphosphine base) ferrocenepalladium(II) (19.5mg, 0.024mmol) and aqueous sodium carbonate (0.30mL, 2M, 0.60mmol). The reaction was heated at 90°C for 12 hours and then cooled to room temperature. The reaction mixture was filtered through a pad of silica gel. The solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 50% ethyl acetate/hexanes) to give the desired coupled product.

Step H: (3S)-1-[(2S,3S)-2-Amino-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1-oxobutyl]-3 - Fluoropyrrolidine trifluoroacetate

To a stirred solution of 61.0 mg of the coupling product from Step G in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. After stirring at room temperature for 1 hour, the solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to afford the title compound. LC/MS 345.0 (M+1).

Example 2

(3S)-1-[(2S,3S)-2-amino-3-(3'-carboxy-1,1'-biphenyl-4-yl)-1-oxobutyl]-3-fluoropyridine Loridine trifluoroacetate

Step A: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-(3'-benzyloxycarbonyl-1,1'- Biphenyl-4-yl)-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Example 1, Step F (214.9 mg, 0.50 mmol) in 5 mL of toluene and 5 mL of ethanol was added 3-benzyloxycarbonylphenylboronic acid (512.4 mg, 2.00 mmol), 1,1 chloride '-Bis(diphenylphosphino)ferrocenepalladium(II) (81.7 mg, 0.100 mmol) and 2M aqueous sodium carbonate (1.25 mL, 2.50 mmol). The reaction was heated at 9OC for 12 hours, then cooled to room temperature. The reaction mixture was filtered through a pad of silica gel. The solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 50% ethyl acetate/hexanes) to give the desired coupled product.

Step B: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-(3'-carboxy-1,1'-biphenyl-4- Base)-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product of Step A in 3 mL of ethyl acetate was added 50 mg of 10% Pd-C. The reaction flask was flushed with nitrogen, then stirred under a hydrogen atmosphere (1 atm) for 12 hours. After the reaction was complete, the solution was passed through a pad of celite. The solvent was removed in vacuo to give the title compound.

Step C: (3S)-1-[(2S,3S)-2-amino-3-(3'-carboxy-1,1'-biphenyl-4-yl)-1-oxobutane base]-3-fluoropyrrolidine trifluoroacetate

The product from Step B was dissolved in 5 mL of dichloromethane and 1 mL of trifluoroacetic acid. The reaction was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to afford the title compound. LC/MS 371.1 (M+1).

Example 3

(3S)-1-[(2SL,3S)-2-amino-3-[3'-(tetrazol-5-yl)-1,1'-biphenyl-4-yl]-1-oxobutane base]-3- Fluoropyrrolidine trifluoroacetate

Step A: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-(3'-cyano-1,1'-biphenyl-4- Base)-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Example 1, Step F (210.0 mg, 0.49 mmol) in 4 mL of toluene and 4 mL of ethanol was added 3-cyanophenylboronic acid (275.6 mg, 1.87 mmol), 1,1'- Bis(diphenylphosphino)ferrocenepalladium(II) (76.0 mg, 93.1 mmol) and 2M aqueous sodium carbonate (1.2 mL, 24.0 mmol). The reaction was heated at 90°C for 12 hours and then cooled to room temperature. The reaction mixture was filtered through a pad of silica gel. The solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 50% ethyl acetate/hexanes) to give the desired coupled product.

Step B: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-[3'-(tetrazol-5-yl)-1,1'- Biphenyl-4-yl]-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Step A (101.0 mg, 0.22 mmol) in 5 mL of toluene was added trimethyltin azide (0.586 mg, 2.85 mmol). The reaction was heated at 100°C for 12 hours and then cooled to room temperature. The solvent was removed and the residue was purified by preparative TLC (silica gel, hexane/ethyl acetate) to give the desired product.

Step C: (3S)-1-[(2S,3S)-2-amino-3-[3'-(tetrazol-5-yl)-1,1'-biphenyl-4-yl]-1- oxygen Butyl]-3-fluoropyrrolidine trifluoroacetate

The product from Step B (56.2 mg) was dissolved in 5 mL of dichloromethane and 1 mL of trifluoroacetic acid and stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to afford the title compound. LC/MS 395.2 (M+1).

Example 4

(3S)-1-[(2S,3S)-2-amino-3-[3'-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl) -1,1'- Biphenyl-4-yl]-1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

Step A: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[3′-(5-oxo-4,5-dihydro- 1,2,4-oxadiazol-3-yl)-1,1'-biphenyl-4-yl]-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Example 3, Step A (140 mg, 0.31 mmol) in 3 mL of ethanol was added 3 mL of hydroxylamine (50% in water). The reaction was heated at 90°C for 12 hours and then cooled to room temperature. The solvent was removed in vacuo and the residue was azeotropically dried with toluene. To a stirred solution of the above residue in 5 mL of dichloromethane was added triethylamine (0.31 mL, 2.22 mmol) followed by ethyl chloroformate (0.155 mL, 1.48 mmol). The reaction was stirred at room temperature for 2 hours before being quenched with saturated aqueous ammonium chloride. The aqueous solution was extracted with ethyl acetate. The combined organic layers were washed with saturated brine, dried over sodium sulfate, and concentrated in vacuo. The residue was dissolved in 5 mL of toluene and heated at 120°C overnight. The solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 25% ethyl acetate/hexanes) to give the product. LC/MS 511.0 (M+1).

Step B: (3S)-1-[(2S,3S)-2-Amino-3-[3'-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3 - Base)-1,1'-biphenyl-4-yl]-1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

To a stirred solution of the product of Step A in 5 mL of dichloromethane was added 1 mL of trifluoroacetic acid. The reaction mixture was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to afford the title compound. LC/MS 411.1 (M+1).

Example 5

(3S)-1-[(2S,3S)-2-amino-3-[4-(6-oxo-1,6-dihydropyridin-3-yl)phenyl]-1-oxo hydrochloride Butyl]-3-fluoropyrrolidine

Step A: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[4-(6-methoxypyridine-3- )phenyl]-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Example 1, Step F (51.0 mg, 0.12 mmol) in 2 mL of toluene and 2 mL of ethanol was added 2-methoxy-5-pyridineboronic acid (71.3 mg, 0.466 mmol), 1 chloride, 1'-bis(diphenylphosphino)ferrocenepalladium(II) (19.5 mg, 0.238 mmol) and 2M aqueous sodium carbonate (0.30 mL, 0.60 mmol). The reaction was heated at 90°C for 12 hours and then cooled to room temperature. The reaction mixture was filtered through a pad of silica gel. The solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 50% ethyl acetate/hexanes) to give the desired coupled product.

Step B: (3S)-1-[(2S,3S)-2-amino-3-[4-(6-oxo-1,6-dihydropyridin-3-yl) hydrochloride Phenyl]-1-oxobutyl]-3-fluoropyrrolidine

The product from Step A was dissolved in 3 mL of concentrated hydrochloric acid (37%). The reaction mixture was heated at 100°C for 48 hours and then cooled to room temperature. The water was removed azeotropically with toluene. Purification by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% concentrated hydrochloric acid) afforded the title compound. LC/MS 344.1 (M+1).

Example 6

(3S)-1-[(2S,3S)-2-amino-3-[4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl]-1 - Oxobutyl]-3-fluoropyrrolidine trifluoroacetate

Step A: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[4-(6-oxo-1,6-dihydro Pyridin-3-yl)phenyl]-1-oxobutyl]-3-fluoropyrrolidine

Example 5, (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[4-(6-methoxypyridin-3-yl)benzene Base]-1-oxobutyl]-3-fluoropyrrolidine (0.65g, 1.42mmol) was mixed with pure pyridine hydrochloride (2.25g, 19.5mmol) and placed in a preheated oil bath (160°C). After stirring at 160°C for 20 minutes, the reaction was cooled to room temperature and neutralized to pH 7 with saturated sodium bicarbonate solution. To this was added 15 mL of 1,4-dioxane followed by di-tert-butyl dicarbonate (2.14 g, 9.82 mmol). After stirring at room temperature for 12 hours, the reaction was partitioned between ethyl acetate and saturated brine. The organic layer was separated and the aqueous layer was extracted with two portions of ethyl acetate. The combined organic layers were dried over sodium sulfate and the solvent was removed by evaporation under reduced pressure. The residue was purified by preparative TLC (silica gel, 50% ethyl acetate/hexanes) to give the desired product.

Step B: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[4-(1-methyl-6-oxo-1,6 Dihydropyridin-3-yl)phenyl]-1-oxobutyl]-3-fluoropyrrolidine

To a stirred solution of the product from Step A (430.0 mg, 0.97 mmol) in 3 mL of DMF was added cesium carbonate (0.57 mg, 1.74 mmol) followed by iodomethane (0.4 mL, 6.4 mmol) at room temperature. After 30 minutes, the reaction mixture was diluted with ethyl acetate, washed with saturated brine, concentrated in vacuo, and the residue was purified by preparative TLC (silica gel, 6% methanol in dichloromethane) to give the product.

Step C: (3S)-1-[(2S,3S)-2-amino-3-[4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl) Phenyl]-1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

The product from Step B was dissolved in 5 mL of dichloromethane and 1 mL of trifluoroacetic acid. After 1 hour of reaction at room temperature, the solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to give the desired product. LC/MS 358.2 (M+1).

Example 7

(3S)-1-[(2S,3S)-2-amino-4-cyclopropyl-3-[4-(1-methyl-6-oxo-1,6-dihydropyridine-3- Base) phenyl] -1-oxobutyl] -3-fluoropyrrolidine trifluoroacetate

Step A: (4R)-3-[(3R)-3-(4-Bromophenyl)hex-5-enoyl]-4-phenyl-1,3-oxazolidine-2- ketone

A 4.5 g portion of the product from Example 1, Step A was converted to the title compound essentially following the procedure outlined in Example 1, Step B, using allylmagnesium bromide.

Step B: (4R)-3-[(3R)-3-(4-Bromophenyl)-4-cyclopropylbutyryl]-4-phenyl-1,3-oxazole alkan-2-one

To a stirred solution of the product of Step A (1.32 g, 3.18 mmol) in 20 mL of diethyl ether was added successively excess diazomethane in diethyl ether and palladium(II) acetate (0.215 g, 0.957 mmol) at 0° C., and The reaction was stirred at 0°C for 2 hours. Excess diazomethane was then quenched with acetic acid. The reaction mixture was filtered through a pad of silica gel and the solvent was removed in vacuo. NMR indicated a 1:1 mixture of desired product and starting material. The same reaction sequence was repeated one more time and the desired product was isolated by flash chromatography (silica gel, 25% ethyl acetate/hexane). LC/MS 430.0 (M+1).

Step C: (4R)-3-[(2R,3S)-2-Bromo-3-(4-bromophenyl)-4-cyclopropylbutyryl]-4-phenyl- 1,3-oxazolidin-2-one

Following essentially the procedure outlined in Example 1, Step C, a 1.16 g portion of the product from Step B was converted to the title compound.

Step D: (4R)-3-[(2S,3S)-2-Azido-3-(4-bromophenyl)-4-cyclopropylbutyryl]-4-benzene Base-1,3-oxazolidin-2-one

Following essentially the procedure outlined in Example 1, Step D, a 0.61 g portion of the product from Step C was converted to the title compound.

Step E: (2S,3S)-2-Azido-3-(4-bromophenyl)-4-cyclopropylbutanoic acid

Following essentially the procedure outlined in Example 1, Step E, a 0.51 g portion of the product from Step D was converted to the title compound.

Step F: (3S)-1-[(2S,3S)-2-Azido-3-(4-bromophenyl)-4-cyclopropylbutanoyl]-3-fluoro pyrrolidine

Following essentially the procedure outlined in Example 1, Step F, a 0.267 g portion of the product from Step E was converted to the title compound.

Step G: (3S)-1-[(2S,3S)-3-(4-Bromophenyl)-2-(tert-butoxycarbonylamino)-4-cyclopropyl Butyryl]-3-fluoropyrrolidine

Following essentially the procedure outlined in Example 1, Step G, a 0.31 g portion of the product from Step F was converted to the title compound.

Step H: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-4-cyclopropyl-3-[4-(6-methoxy ylpyridin-3-yl)phenyl]butyryl]-3-fluoropyrrolidine

Following essentially the procedure outlined in Example 5, Step A, a 0.17 g portion of the product from Step G was converted to the title compound. LC/MS 498.1 (M+1).

Step I: (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-4-cyclopropyl-3-[4-(6-oxo- 1,6-Dihydropyridin-3-yl)phenyl]butyryl]-3-fluoropyrrolidine

The product from Step H (0.18 g, 0.361 mmol) was mixed with neat pyridine hydrochloride (0.83 g, 7.21 mmol) and placed in a preheated oil bath (160°C). After stirring at 160°C for 20 minutes, the reaction was cooled to room temperature and neutralized to pH 7 with saturated aqueous sodium bicarbonate. Then 10 mL of 1,4-dioxane was added followed by di-tert-butyl dicarbonate (0.473 g, 2.17 mmol). After stirring at room temperature for 12 hours, the reaction was partitioned between ethyl acetate and saturated brine. The organic layer was separated and the aqueous layer was extracted with two portions of ethyl acetate. The combined organic layers were dried over sodium sulfate and the solvent was removed by evaporation under reduced pressure. Purification by preparative TLC (silica gel, 50% ethyl acetate/hexanes) afforded the desired product.

Step J: (3S)-1-[(2S,3S)-2-amino-4-cyclopropyl-3-[4-(1-methyl-6-oxo-1,6-dihydro Pyridin-3-yl)phenyl]-1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

Following essentially the procedure outlined in Example 6, Steps B and C, a 0.056 g portion of the product from Step I was converted to the title compound. LC/MS 398.3 (M+1).

Example 8

(3S)-1-[(2S,3S)-2-amino-3-[4-(5-bromo-6-oxo-1,6-dihydropyridin-3-yl)phenyl]-1- oxygen Butyl]-3-fluoropyrrolidine trifluoroacetate

To (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-[4-(6-oxo-1,6-dihydropyridin-3-yl)phenyl ]-1-oxobutyl]-3-fluoropyrrolidine (Example 6, Step A, 111.4 mg, 0.251 mmol) was added to a stirred solution of pyridinium tribromide (0.098 g, 0.306 mmol). After 2 hours at room temperature, the solvent was removed in vacuo and the residue was purified by preparative TLC (silica gel, 9% methanol/dichloromethane) to give 47 mg of product which was dissolved in 5 mL of dichloromethane and 1 mL of trifluoroacetic acid. After 1 hour of reaction at room temperature, the solvent was removed in vacuo and the residue was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA) to afford the title compound. LC/MS 424.0 (M+1).

Example 9

(3S)-1-[(2S,3S)-2-amino-3-[3'-[(tert-butylamino)carbonyl-1,1'-biphenyl-4-yl]-1-oxo Butyl]-3-fluoropyrrolidine trifluoroacetate

To (3S)-1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-(3'-carboxy-1,1'-biphenyl-4-yl)-1-oxo Butyl]-3-fluoropyrrolidine (Example 2, Step B) (37.6 mg, 0.08 mmol) was added to a stirred solution of hydrochloric acid 1-( 3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC, 23mg, 0.12mmol), hydroxybenzotriazole (HOBT, 16.2mg, 0.12mmol) and tert-butylamine (0.32mL, 0.16 mmol). After stirring at room temperature for 16 hours, the reaction was diluted with ethyl acetate. The organic phase was washed successively with 0.5N hydrochloric acid, saturated aqueous sodium bicarbonate, water and saturated brine, dried (magnesium sulfate), and concentrated under reduced pressure to obtain the crude coupling product, which was prepared by preparative thin-layer chromatography (silica gel, 50% acetic acid ethyl ester/hexane eluent). The resulting product was dissolved in 1 mL of dichloromethane and treated with 1 mL of trifluoroacetic acid. The reaction was stirred at room temperature for 1 hour, then concentrated under reduced pressure to afford the title compound. LC/MS 426.3 (M+1).

Example 10

(3S)-1-[(2S,3S)-2-amino-3-[3'-[[(trifluoromethyl)sulfonyl]amino]-1,1'-biphenyl-4-yl]- 1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

Step A: 3-Iodo-phenyl-trifluoromethanesulfonamide

To a solution of 3-iodoaniline (0.36 mL, 3.0 mmol) in 15 mL of dichloromethane was added trifluoroacetic anhydride (1.00 mL, 6.0 mmol) dropwise. Pyridine (1.21 mL, 15.0 mmol) was added and the resulting clear solution was stirred at room temperature for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (10 mL), and 1N aqueous sodium hydroxide solution (10 mL) was added. After stirring at room temperature for 30 minutes, the two layers were separated. The organic phase was washed with saturated brine, dried (magnesium sulfate), and concentrated. The crude product was purified by flash chromatography (silica gel, 30% ethyl acetate-hexane eluent) to give the desired product as a colorless oil.

Step B: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-[4-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)phenyl]-1-oxobutyl]-3-fluoropyrrolidine

To a thick-walled sealable test tube was added the (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-(4-bromophenyl from Example 1, Step F )-1-oxobutyl]-3-fluoropyrrolidine (1.0g, 2.33mmol), bis(pinacolato)diboron (1.78g, 6.99mmol), potassium acetate (1.14g, 11.66 mmol) and 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (0.38 g, 0.466 mmol). Dimethyl sulfoxide (15 mL) was added, and the tube was flushed with nitrogen and sealed. The reaction mixture was warmed at 80°C overnight, cooled to room temperature and partitioned between water and ethyl acetate. The mixture was extracted with two more portions of ethyl acetate. The combined ethyl acetate extracts were washed with saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure to give a brown oil. Purification by flash chromatography (silica gel, 5-25% ethyl acetate/hexanes gradient) afforded the desired product as a foam. LC/MS 477.2 (M+1).

Step C: (3S)-1-[(2S,3S)-2-amino-3-[3'-[[(trifluoromethyl)sulfonyl]amino]-1,1'-link Benzene-4-yl]-1-oxobutyl]-3-fluoropyrrolidine trifluoroacetate

To a stirred solution of the product from Step A (70.2 mg, 0.2 mmol) in 0.75 mL of ethylene glycol dimethyl ether and 0.75 mL of water was added 47.6 mg (0.10 mmol) of the product from Step B, 1,1'-bis( Diphenylphosphino)ferrocenepalladium(II) (16.4 mg, 0.02 mmol) and potassium phosphate (63.6 mg, 0.3 mmol). The reaction mixture was heated at 90°C for 16 hours. After cooling to room temperature, the mixture was diluted with ethyl acetate, washed successively with saturated aqueous sodium bicarbonate solution, water and saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure to obtain the crude coupling product, which was subjected to preparative thin-layer chromatography. (silica gel, ethyl acetate eluent). The resulting coupled product was further purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water containing 0.1% TFA). The purified product was dissolved in 0.5 mL of dichloromethane and treated with 0.5 mL of trifluoroacetic acid. After stirring at room temperature for 1 hour, the solvent was removed in vacuo and the residue was purified by preparative HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water with 0.1% TFA) to afford the title compound. LC/MS 474.2 (M+1).

Example 11

(3S)-1-[(2S,3S)-2-Amino-3-[4-([1,2,4]triazolo[4,3-α]pyridin-6-yl)phenyl]- 1-oxo Butyl]-3-fluoropyrrolidine

Step A: 5-iodo-2-hydrazinopyridine

A mixture of 2-chloro-5-iodopyridine (1.918 g, 8.01 mmol), anhydrous hydrazine (1.26 mL, 40.05 mmol) and pyridine (30 mL) was warmed at reflux for 18 hours. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was partitioned between dichloromethane and 1N aqueous sodium hydroxide solution. The organic layer was separated, dried (magnesium sulfate) and concentrated under reduced pressure. The residue was triturated with hexanes and the resulting precipitate collected and dried in vacuo to give the title compound as off-white crystals. LC/MS 235.8 (M+1).

Step B: 6-iodo-[1,2,4]triazolo[4,3-α]pyridine

A mixture of the product from Step A above (2.0 g, 8.51 mmol) in triethyl orthoformate (100 mL) was warmed at reflux for 18 hours. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in 250 mL of dichloromethane and filtered through a pad of silica gel. The compound was eluted from the silica gel by washing the pad with 20% methanol/dichloromethane, and the filtrate was concentrated to dryness, then redissolved in dichloromethane. Hexane was added to give a precipitate which was collected. The filtrate was concentrated to half volume then diluted with additional hexanes to give a second crop of product. The combined solids were dried in vacuo to give the title compound as a light yellow solid. LC/MS 246.0 (M+1).

Step C: (3S)-1-[(2S,3S)-2-amino-3-[4-([1,2,4]triazolo[4,3-α]pyridin-6-yl)benzene Base]-1-oxobutyl]-3-fluoropyrrolidine

[ (2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane Alk-2-yl)phenyl]-1-oxobutyl]-3-fluoropyrrolidine, 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (21.0mg , 0.26mmol) and potassium phosphate (83mg, 0.39mmol). The reaction mixture was heated at 90°C for 16 hours and then cooled to room temperature. The mixture was diluted with ethyl acetate, the solution was washed successively with water and saturated brine, dried (magnesium sulfate) and concentrated in vacuo under reduced pressure to obtain the crude coupling product, which was purified by preparative TLC (silica gel, 5% methanol/ dichloromethane eluent). The resulting product was dissolved in 1 mL of dichloromethane and treated with 1 mL of trifluoroacetic acid. The reaction was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified by preparative thin layer chromatography (silica gel, 10% methanol/1% ammonium hydroxide/dichloromethane eluent) to afford the title compound. LC/MS 368.2 (M+1).

Example 12

(3S)-1-[(2S,3S)-2-amino-3-carboxy-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1-oxopropyl]- 3- Fluoropyrrolidine trifluoroacetate

Step A: trans-4-(4-bromophenyl)-3-buten-2-one

To 25.0 g (110 mmol) of 4-bromocinnamic acid dissolved in anhydrous dichloromethane (500 mL) was added EDC (28.8 g, 150 mmol), HOBT (20.3 g, 150 mmol), N,O-dimethylhydroxylamine hydrochloride (14.6 g, 150 mmol) and N,N'-diisopropylethylamine (23 mL, 150 mmol). After stirring at room temperature for 24 hours, the reaction was concentrated and then diluted with 400 mL of 10% aqueous hydrochloric acid. The resulting mixture was then extracted with three 300-mL portions of ether, and the combined organic phases were washed sequentially with 10% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (100 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo to give the Weinreb amide as a viscous oil which was used without further purification. To this oil was added 300 mL of anhydrous tetrahydrofuran and the resulting solution was cooled to -78°C. To this solution was added 60 mL of methylmagnesium bromide (180 mmol, 3N in ether). The stirred mixture was slowly warmed to 0°C over 1 hour. The mixture was then carefully quenched with water and 5% aqueous hydrochloric acid (100 mL each), then concentrated to remove tetrahydrofuran. The resulting mixture was extracted with three 300-mL portions of ether, and the combined organic phases were washed sequentially with 5% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (100 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo to give a viscous oil. The crude product was purified by flash chromatography (silica gel, 0-15% ethyl acetate/hexanes gradient) on a Biotage® system to afford the title compound as a pale yellow crystalline solid. LC/MS 225.0 (M+1), 227.0 (M+3).

Step B: (2S,3E)-4-(4-Bromophenyl)-3-buten-2-ol

To 5.55 g (24.7 mmol) of the ketone from step A dissolved in 100 mL of toluene was added 3.7 mL (3.7 mmol, 1M in toluene) of (R)-2-methyl-CBS-oxazaborole alkane catalyst and the resulting mixture was stirred at ambient temperature for 15 minutes. The mixture was cooled to -78°C and 4.0 mL (37.1 mmol) catecholborane in 30 mL toluene was added dropwise over 30 minutes. After the addition, the slurry was stirred at -78°C for 60 minutes, during which time it slowly became homogeneous. The solution was then stirred at -78°C for an additional 4 hours (reaction time varied from 4 to 24 hours) until TLC showed complete disappearance of the starting material. Next, the reaction mixture was diluted with 100 mL of water, and the resulting mixture was extracted with three 100-mL portions of ether. The organic phases were then combined, washed with two 100-mL portions of 1 N aqueous NaOH solution, two 100-mL portions of 5% hydrochloric acid solution, multiple 100-mL portions of saturated brine, dried over magnesium sulfate, filtered, and evaporated in vacuo to give the crude waxy solid . The crude product was then purified by flash chromatography (silica gel, 0-20% ethyl acetate/hexane gradient) on a Biotage® system to afford the alcohol as a colorless crystalline solid. The compound was recrystallized from hexane to give the alcohol as colorless crystals (96% ee by Mosher ester analysis). LC/MS 209.0 ( _MH2O +1), 211.0 ( _MH2O +3).

Step C: N-(tert-butoxycarbonyl)glycine (1S,2E)-3-(4-bromophenyl)-1-methylprop-2-ene ester

To 12.6 g (55 mmol) of the alcohol from step B dissolved in dry dichloromethane (300 mL) was added EDC (23 g, 120 mmol), HOBT (16 g, 120 mmol), N-(tert-butoxycarbonyl)glycine (21 g , 120mmol) and N,N'-diisopropylethylamine (19mL, 120mmol). After 5 hours, the mixture was concentrated and diluted with 200 mL of 10% aqueous hydrochloric acid. The resulting mixture was then extracted with three 300-mL portions of diethyl ether, and the combined organic phases were washed sequentially with 5% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (100 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo. The crude product was obtained as a viscous oil. The crude product was purified by flash chromatography (silica gel, 0-20% ethyl acetate/hexanes gradient) on a Biotage® system to afford the title compound as a colorless crystalline solid. LC/MS 328.1 (M-tert-butyl+1), 330.1 (M-tert-butyl+3).

Step D: (βS)-4-Bromo-N-(tert-butoxycarbonyl)-β-[(1E)-prop-1-enyl]-L-phenylalanine methyl ester

The ester of Step C (18.1 g, 47 mmol) in anhydrous THF (50 mL) was added via cannula to 105 mL (105 mmol, 1 M in THF) of hexamethyldisilylazide precooled to -78 °C. Lithium nitride (lithium hexamethyldisilazide) solution. After stirring at this temperature for 10 minutes, 55 mL of zinc chloride solution (55 mmol, 1M in ether) was added at -78°C. The resulting mixture was stirred at -78°C for 5 hours, then allowed to warm slowly to room temperature over 3 hours. After stirring at room temperature for an additional 2 hours, the mixture was quenched with water and 5% hydrochloric acid (100 mL each). The resulting mixture was then extracted with three 300-mL portions of ethyl acetate, and the combined organic phases were washed sequentially with 5% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (200 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo to give the crude product as a yellow foam. LC/MS 384.1(M+1), 386.1(M+3). The crude product was dissolved in 500 mL of 1:1 ether/methanol and cooled to 0 °C. A solution of trimethylsilyldiazomethane (75 mL, 150 mmol, 2M in hexanes) was added in portions until yellow color persisted. After warming to room temperature, the solution was stirred for an additional 8 hours, then concentrated in vacuo. The crude product was purified by flash chromatography (silica gel, 0-15% ethyl acetate/hexanes gradient) on a Biotage® system to afford the title compound as a colorless oil. LC/MS 298.0 (M-Boc+1), 300.0 (M-Boc+3).

Step E: (2S,3S,4E)-2-[(tert-butoxycarbonyl)amino]-3-(4'-fluoro-1,1'-biphenyl-4-yl)-4- Methyl hexenoate

To a stirred solution of the product from Step D (5.5 g, 13.8 mmol) in 40 mL of toluene and 7 mL of 2M aqueous sodium carbonate (14 mmol) was added 4-fluorophenylboronic acid (2.52 g, 18 mmol) and tetrakis(triphenylphosphino ) Palladium(0) (2 g, 1.7 mmol). The reaction was heated at 140°C for 20 hours, then cooled to room temperature and diluted with 100 mL of water. The resulting mixture was then extracted with three 150-mL portions of diethyl ether, and the combined organic phases were washed sequentially with 5% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (100 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo to give the crude product as a viscous oil. The crude product was purified by flash chromatography (silica gel, 0-15% ethyl acetate/hexanes gradient) on a Biotage® system to afford the title compound as a colorless waxy solid. LC/MS 414.3 (M+1).

Step F: (2S,3S,4E)-2-[(tert-butoxycarbonyl)amino]-3-(4'-fluoro-1,1'-biphenyl-4-yl)-4- hexenoic acid

A solution of the product from Step E (3.55 g, 8.9 mmol) in 250 mL of 3:1:1 tetrahydrofuran/methanol/1N aqueous lithium hydroxide (50 mL, 50 mmol) was stirred at room temperature for 15 hours, then concentrated and washed with 200 mL of 10% hydrochloric acid Acidification of the aqueous solution. The resulting mixture was then extracted with three 150-mL portions of ethyl acetate, and the combined organic phases were washed successively with 5% hydrochloric acid and saturated brine (100 mL each). The organic phase was dried over magnesium sulfate, filtered and evaporated in vacuo to give the acid as a colorless foamy solid which was used without further purification. LC/MS 385.2 (M+1).

Step G: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-(4'-fluoro-1,1'-biphenyl-4- Base)-1-oxohex-4-enyl]-3-fluoropyrrolidine

To 2.11 g (5.49 mmol) of the acid from Step F dissolved in dry dichloromethane (100 mL) was added EDC (1.34 g, 7.0 mmol), HOBT (0.95 g, 7.0 mmol), hydrochloric acid (3S)-3- Fluoropyrrolidine (880 mg, 7.0 mmol) and N,N'-diisopropylethylamine (1.1 mL, 7.0 mmol). After stirring at room temperature for 48 hours, the reaction mixture was concentrated and diluted with 100 mL of 10% aqueous hydrochloric acid. The resulting mixture was then extracted with three 150-mL portions of ethyl acetate, and the combined organic phases were washed sequentially with 5% hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine (100 mL each). The organic phase was then dried over magnesium sulfate, filtered and evaporated in vacuo to give the crude product as a viscous oil. The crude product was purified by flash chromatography (silica gel, 0-40% ethyl acetate/hexanes gradient) on a Biotage® system to afford the title compound as a colorless solid. LC/MS 471.3 (M+1).

Step H: (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-carboxy-3-(4'-fluoro-1,1'- Biphenyl-4-yl)-1-oxopropyl]-3-fluoropyrrolidine

A solution of 1.0 g (2.12 mmol) of the olefin from Step G in 1:1 methanol/dichloromethane (150 mL) was flushed with oxygen for 2 minutes and then cooled to -78°C. Ozone was bubbled through the solution until a light blue color persisted (about 5 minutes), then oxygen was bubbled in until the blue color disappeared again. Dimethyl sulfide (2 mL, excess) was added and the resulting mixture was allowed to warm to room temperature and stirred for an additional 20 minutes. The reaction mixture was concentrated, diluted with 200 mL of ethyl acetate, and washed successively with 5% hydrochloric acid, saturated aqueous sodium bicarbonate solution and saturated brine (50 mL each). The organic phase was dried over magnesium sulfate, filtered and evaporated in vacuo to give the crude aldehyde. Crude aldehyde, sodium dihydrogen phosphate (439 mg, 3.18 mmol), sodium chlorite (580 mg, 6.4 mmol), and isobutene (4.25 mL, 8.5 mmol, 2M in THF) were dissolved in 50 mL of 4:1 tert-butanol/ Stir in water for 8 hours, then concentrate. The crude mixture was diluted with 50 mL of 5% aqueous hydrochloric acid and extracted with three 150-mL portions of ethyl acetate. The combined organic phases were washed successively with 5% hydrochloric acid and saturated brine (50 mL each), dried over sodium sulfate, filtered and evaporated in vacuo to give the crude product as a colorless crystalline solid. The crude product was purified by HPLC (YMC Pro-C18 column, gradient elution, 30-95% acetonitrile/water with 0.1% TFA) to afford the title compound as a colorless crystalline solid. LC/MS 475.2 (M+1).

Step I: (3S)-1-[(2S,3S)-2-amino-3-carboxy-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1-oxo Propyl]-3-fluoropyrrolidine trifluoroacetate

The acid from Step H (55 mg, 0.12 mmol) was dissolved in 20 mL of 1:1 trifluoroacetic acid/dichloromethane and stirred for 60 minutes, then concentrated. The crude product was purified by reverse phase HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water with 0.1% TFA) to afford the title compound as a colorless crystalline solid. LC/MS 375.2 (M+1).

Example 13

(3S)-1-[(2S,3S)-2-amino-3-(dimethylaminocarbonyl)-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1- Oxopropyl]-3-fluoropyrrolidine trifluoroacetate

To 200 mg (0.42 mmol) of Example 12, step H, dissolved in dry dichloromethane (10 mL) (3S)-1-[(2S,3S)-2-[(tert-butoxycarbonyl)amino] -3-carboxy-3-(4'-fluoro-1,1'-biphenyl-4-yl)-1-oxopropyl]-3-fluoropyrrolidine was added with EDC (116mg, 0.6mmol), HOBT (81 mg, 0.6 mmol), dimethylamine (0.4 mL, 0.8 mmol, 2.0 M in THF) and N,N'-diisopropylethylamine (0.091 mL, 0.6 mmol). After stirring at room temperature for 48 hours, the reactant was diluted with 200 mL of ethyl acetate, and then washed successively with 5% hydrochloric acid, saturated aqueous sodium bicarbonate solution and saturated brine (50 mL each time). The organic phase was dried over magnesium sulfate, filtered and evaporated in vacuo to give the amide as a yellow foam. This material was dissolved in 50 mL of 1:1 trifluoroacetic acid/dichloromethane and stirred for 60 minutes, then concentrated. The crude product was purified by HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water with 0.1% TFA) to afford the title compound as a pale yellow crystalline solid. LC/MS 402.3 (M+1).

Example 14

1-[(2S,3S)-2-amino-3-(dimethylaminocarbonyl)-3-(4-[1,2,4]triazolo[1,5-α] hydrochloride Pyridin-6-ylphenyl)-1-oxopropyl]-3,3-difluoropyrrolidine

Step A: 1-[(2S,3S,4E)-3-(4-Bromophenyl)-2-(tert-butoxycarbonylamino)hex-4-enoyl Base]-3,3-difluoropyrrolidine

To 25g (62.8mmol) of (βS)-4-bromo-N-(tert-butoxycarbonyl)-β-[(1E)-prop-1-enyl]-L-phenylalanine methyl ester (implementation Example 12, Step D) To a solution in 600 mL of tetrahydrofuran (THF), 200 mL of methanol and 200 mL (200 mmol) of 1N aqueous sodium hydroxide were added successively. The reaction mixture was stirred at ambient temperature for 3 hours, then methanol and THF were removed under reduced pressure. To the aqueous mixture was added 250 mL of 1N hydrochloric acid and the mixture was extracted with ethyl acetate (3 x 300 mL). The combined organic extracts were washed with brine (300 mL), then dried over sodium sulfate, filtered, and concentrated in vacuo to afford the carboxylic acid, which was used without further purification.

The above acid was mixed with 18.6 g (130 mmol) of 3,3-difluoropyrrolidine, 17.5 g (130 mmol) of HOBt, 22.8 mL (130 mmol) of N,N-diisopropylethylamine and 300 mL of DMF. Then 25 g (130 mmol) of EDC were added and the solution was stirred at ambient temperature for 12 hours under a nitrogen atmosphere. Ethyl acetate (1.5 L) was added and the mixture was washed with 0.5N aqueous sodium bicarbonate (3 x 400 mL), 1N hydrochloric acid (2 x 400 mL) and brine (400 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford the title compound , which was pure enough for subsequent steps. MS 375.1 (M+1-Boc).

Step B: 1-[(2S,3S)-3-(4-Bromophenyl)-2-(tert-butoxycarbonylamino)-3-(dimethylamino ylcarbonyl)-1-oxopropyl]-3,3-difluoropyrrolidine

1.5 L of water was added to the round bottom flask, and 80 g (374 mmol) of sodium periodate were added. The mixture was stirred until homogeneous, and then 1.2 g (7.5 mmol) of potassium permanganate was added to the mixture. To this deep purple solution was added 5.7 g (41.1 mmol) of potassium carbonate powder (-325 mesh) and 17.7 g (37.4 mmol) of the product from Step A in 500 mL of tert-butanol. The reaction mixture was stirred at ambient temperature for 24 hours, then treated with 50 mL of saturated aqueous sodium sulfite, acidified with 1N aqueous hydrochloric acid (400 mL), and extracted with ethyl acetate (3 x 400 mL). The combined organic extracts were washed with brine (3 x 400 mL), and the resulting clear solution was dried over sodium sulfate, filtered, and concentrated in vacuo to give the crude acid, which was used without further purification.

The above acid was mixed with 10.1 g (74.8 mmol) of HOBt and 300 mL of DMF. Then 13 mL (74.8 mmol) of N,N-diisopropylethylamine, 37.4 mL (74.8 mmol) of 2N dimethylamine in THF and 14.3 g (74.8 mmol) of EDC were added successively to the solution. The reaction mixture was then stirred at ambient temperature for 12 hours. Ethyl acetate (1.2 L) was then added and the mixture was washed with 0.5N aqueous sodium bicarbonate (3 x 400 mL), 1N aqueous hydrochloric acid (2 x 300 mL) and brine (400 mL), dried over sodium sulfate, filtered and concentrated in vacuo. Purification by Biotage Horizon® system flash chromatography (silica gel, 1:1 ethyl acetate/hexane-100% ethyl acetate-10% methanol/ethyl acetate gradient) afforded the title compound. MS 450.0 (M+1-tert-butyl).

Step C: 1-[(2S,3S)-2-(tert-butoxycarbonylamino)-3-(dimethylaminocarbonyl)-1-oxo -3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propyl]-3,3-di Fluoropyrrolidine

To 11.5 g (22.8 mmol) of the bromide from Step B was added 11.5 g (45.6 mmol) of pinacol diborate, 3.7 g (4.6 mmol) of [1,1'-bis(diphenylphosphino) dichloride Ferrocene]palladium(II) (complex with dichloromethane (1:1)), 11.2 g (114 mmol) potassium acetate and 70 mL dimethylsulfoxide (DMSO). Nitrogen was then bubbled through the mixture for 3 minutes, and the mixture was stirred at 80° C. for 4 hours under nitrogen. The mixture was allowed to cool to ambient temperature, then filtered through a pad of silica gel, rinsing with excess ethyl acetate. The solution was washed with two portions of brine, dried over sodium sulfate, filtered and concentrated in vacuo. Purification by flash chromatography (silica gel, 40% ethyl acetate/hexane-100% ethyl acetate-20% methanol/ethyl acetate gradient) on a Biotage Horizan® system afforded the title compound. MS 496.3 (M+1-tert-butyl).

Step D: 1-[(2S,3S)-2-amino-3-(dimethylaminocarbonyl)-3-(4-[1,2,4]triazole hydrochloride And[1,5-α]pyridin-6-ylphenyl)-1-oxopropyl]-3,3-difluoropyrrolidine

To 9.27 g (16.8 mmol) of the compound from step C in 200 mL of ethanol/toluene (1:1) was added 6.7 g (33.6 mmol) of 6-bromo[1,2,4]triazolo[1,5-α ]pyridine (intermediate 18), 2.9 g (3.6 mmol) of [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (complex with dichloromethane , 1:1) and 42 mL (84 mmol) of 2N aqueous sodium carbonate. The reaction mixture was stirred at 90°C for 12 hours under nitrogen. After cooling to ambient temperature, 600 mL of ethyl acetate was added to the mixture, and the organic phase was washed successively with 0.5N aqueous sodium bicarbonate solution and brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by reverse phase HPLC (Kromasil C ₈ 16 microns, isocratic, 40% acetonitrile/water with 0.1% TFA) on a Kiloprep-100G system to give the coupled product.

The above intermediate was then dissolved in a 1:1 mixture of dichloromethane and TFA, stirred at room temperature for 30 minutes, then concentrated in vacuo. The product was purified by reverse phase HPLC (Kromasil _C8 16 micron, gradient elution, 0%-65% acetonitrile/water with 0.1% TFA) on a Kiloprep-100G system to give the product as a TFA salt. The salt was then dissolved in water and the aqueous solution was adjusted to pH 2 by the addition of 2N aqueous sodium carbonate. After extracting the aqueous mixture with 3:1 chloroform:isopropanol (5 x 300 mL), the combined organic layers were washed once with brine, then dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting amine was then dissolved in dichloromethane, and 30 mL of 2N hydrogen chloride in ether was added to the solution. After stirring for 60 minutes, the solution was evaporated to give the title compound as a white hydrochloride salt. The compound was further purified by recrystallization (ethanol/ether) and then lyophilized from water/acetonitrile (40:60, 100 mL) to afford the title compound. MS 443.2 (M+1). 500MHz ¹ H NMR (CD ₃ OD) δ9.12(s, 1H), 8.48(s, 1H), 8.06(dd, J=9.4, 1.6Hz, 1H), 7.87(d, J=9.3Hz, 1H) , 7.85 (d, J=8.0Hz, 2H), 7.58 (dd, J=8.2, 1.6Hz, 2H), 4.69 (dd, J=57.4, 8.4Hz, 1H), 4.58 (dd, J=13.8, 8.2 Hz, 1H), 4.52-3.74 (m, 6H), 2.96 (s, 3H), 2.94 (d, J = 1.1 Hz, 3H), 2.69-45 (m, 2H).

Example 15

1-[(2S,3S)-2-amino-3-(dimethylaminocarbonyl)-3-(4-[1,2,4]triazolo[1,5-α]pyridine- 7-ylphenyl)-1-oxopropyl]-3,3-difluoropyrrolidine trifluoroacetate

Under nitrogen atmosphere, the intermediate of Example 14, Step C (48 mg, 0.090 mmol), intermediate 19 (48 mg, 0.18 mmol) and tetrakis(triphenylphosphino)palladium(0) (12 mg) were dissolved in 1.2 mL A mixture of dimethoxyethane, 0.30 mL ethanol, and 0.30 mL 2M aqueous sodium carbonate was warmed at 84°C for 18 hours. The mixture was cooled to room temperature, diluted with 12 mL of ethyl acetate and filtered through a pad of celite. The filtrate was concentrated under reduced pressure and the residue was purified by preparative thin layer chromatography (1 mm silica gel; 12:1 dichloromethane:10% ammonium hydroxide in methanol eluent) to afford 39 mg of the protected intermediate, which was Dissolve in 4 mL of dichloromethane and treat with 2 mL of trifluoroacetic acid. After 1 hour at room temperature, the volatiles were removed under a stream of nitrogen and the residue was triturated with anhydrous ether to give the title compound as a white powder. MS 443.2 (M+1). 500MHz ¹ H NMR (CD ₃ OD) δ8.89(d, J=7.1Hz, 1H), 8.49(s, 1H), 8.05(s, 1H), 7.93(d, J=8.2Hz, 2H), 7.40 (m, 3H), 4.20-4.80 (m, 3H), 3.75-4.05 (m, 3H), 2.98 (s, 3H), 2.95 (s, 3H), 2.55 (m, 2H).

Example 16

1-[(2S,3S)-2-amino-3-(dimethylaminocarbonyl)-3-(4-pyrazolo[1,5-α]pyrimidin-5-ylbenzene base)-1-oxopropyl]-3,3-difluoropyrrolidine trifluoroacetate

Example 14, Intermediate (48 mg, 0.090 mmol) of Step C with Intermediate 20 (38 mg, 0.25 mmol) and tetrakis(triphenylphosphino)palladium(0) was carried out as described for the preparation of Example 15. reaction. The intermediate was purified by preparative thin layer chromatography (1 mm silica gel; 6% methanol in dichloromethane eluent) and deprotected with trifluoroacetic acid as described in Example 15 to afford the title compound as a white solid . MS 443.2(M+1).

Example 17

1-[(2S,3S)-2-amino-3-(methylaminocarbonyl)-3-(4-[1,2,4]triazolo[1,5-α]pyridine-6- phenyl)-1-oxopropyl]-3,3-difluoropyrrolidine trifluoroacetate

Step A: 1-[(2S, 3R and 3S)-2-(tert-butoxycarbonylamino)-3-carboxy-1-oxo-3-[4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propyl]-3,3-difluoropyridine Lolidine

1.5 L of water was added to the round bottom flask and 80 g (374 mmol) of sodium periodate were added. The mixture was stirred until homogeneous, and then 1.2 g (7.5 mmol) of potassium permanganate was added to the mixture. To this dark purple solution was added 5.7 g (41.1 mmol) of potassium carbonate powder (~325 mesh) and 17.7 g (37.4 mmol) of Example 14, Step A, 1-[(2S,3S, 4E)-3-(4-Bromophenyl)-2-(tert-butoxycarbonylamino)hex-4-enoyl]-3,3-difluoropyrrolidine. The reaction mixture was stirred at ambient temperature for 24 hours, then treated with 50 mL of saturated aqueous sodium sulfite, acidified with 1N aqueous hydrochloric acid (400 mL), and extracted with ethyl acetate (3 x 400 mL). The combined organic extracts were washed with brine (3 x 400 mL), and the resulting clear solution was dried over sodium sulfate, filtered, and concentrated in vacuo to give the crude acid, which was used without further purification.

A portion of 6.0g (12.6mmol) of the above acid was mixed with 4.6g (18.0mmol) of diboronic acid pinacol ester, 400mg (0.49mmol) of [1,1'-bis(diphenylphosphino) dichloride Iron]palladium(II) (complex with dichloromethane (1:1)), 7.5 g (76 mmol) of potassium acetate and 40 mL of dimethyl sulfoxide (DMSO) were mixed. After flushing with nitrogen, the mixture was stirred at 100° C. for 10 hours under nitrogen. The mixture was cooled to ambient temperature, acidified with 1N aqueous hydrochloric acid (100 mL), then filtered through a pad of celite, then rinsed with ethyl acetate (200 mL). The layers were separated and the organic layer was extracted with ethyl acetate (2 x 200 mL). The organic layers were then combined, washed with two portions of brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The resulting dark brown solid was pure enough for subsequent steps. MS 425.4 (M+1-Boc).

Step B: 1-[(2S, 3R and 3S)-2-(tert-butoxycarbonylamino)-3-carboxy-3-(4-[1,2,4]tri Azolo[1,5-α]pyridin-6-ylphenyl)-1-oxopropyl]-3,3-difluoropyrrolidine

To the boronic ester of step A in 80 mL of ethanol/toluene (1:1) was added 6.0 g (11.4 mmol) of 6-bromo[1,2,4]triazolo[1,5-α]pyridine ( Intermediate 18), 400 mg (0.49 mmol) of [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (complex with dichloromethane, 1:1 ) and 37 mL (74 mmol) of 2N aqueous sodium carbonate. The reaction mixture was stirred at 100°C for 18 hours under nitrogen. After cooling to ambient temperature, 600 mL of ethyl acetate was added to the mixture, and the organic phase was washed successively with 1N hydrochloric acid and brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by reverse phase HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water with 0.1% TFA) to give the pure coupled product as a mixture of diastereomers at the benzyl position. MS 516.4(M+1).

Step C: 1-[(2S,3S)-2-Amino-3-(methylaminocarbonyl)-3-(4-[1,2,4]triazolo[1,5-α] Pyridin-6-ylphenyl)-1-oxopropyl]-3,3-difluoropyrrolidine trifluoroacetate

To 150 mg (0.30 mmol) of the acid from step B and 92 mg (0.80 mmol) of N-hydroxysuccinimide in 20 mL of dichloromethane was added 160 mg (0.80 mmol) of EDC and the resulting mixture was stirred at ambient temperature under nitrogen. solution for 12 hours. The reaction mixture was quenched with water, then extracted with a 3:1 chloroform/isopropanol (IPA) solution. The combined organic layers were washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo to give the N-hydroxysuccinimide ester. The crude product was dissolved in 10 mL of dioxane and 10 mL of methylamine (2N in THF, 20 mmol) and the mixture was stirred at ambient temperature for 3 hours. The mixture was then concentrated and the residue was purified by reverse phase HPLC (YMC Pro-C18 column, gradient elution, 10-90% acetonitrile/water with 0.1% TFA) to give pure methylamide as a mixture of diastereomers. MS 529.5(M+1).

The above product was then dissolved in a 1:1 mixture of dichloromethane and TFA, stirred at room temperature for 60 minutes, then concentrated in vacuo. This material was then dissolved in 150 mL 3:1 chloroform/IPA and washed with 50 mL saturated aqueous sodium bicarbonate to form the free base. The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting mixture of diastereomers was separated by preparative TLC (10% methanol/dichloromethane) to afford the free base of the title compound as the less polar, faster eluting diastereomer. The free base was again subjected to reverse phase HPLC (YMCPro-C18 column, gradient elution, 0%-50% acetonitrile/water with 0.1% TFA) to afford the title compound. MS 429.4(M+1).

The following intermediates were used in the preparation of some of the compounds of the invention listed in Tables 1-3.

Intermediate 6

6-iodoimidazo[1,2-α]pyridine

Step A: 2-Amino-5-iodopyridine

A mixture of 2-aminopyridine (941 mg, 10 mmol), iodine (980 mg, 3.86 mmol) and periodic acid (547 mg, 2.4 mmol) in 6.6 mL of acetic acid, 2.5 mL of water and 0.16 mL of concentrated sulfuric acid was heated at 80 °C for 2 hours . The reaction was cooled to room temperature and poured into aqueous sodium thiosulfate. The aqueous solution was extracted several times with dichloromethane, the combined extracts were dried (magnesium sulfate) and concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (silica gel, 25% ethyl acetate/hexanes) to give the desired product. LC-MS 220.8 (M+1).

Step B: 6-iodoimidazo[1,2-α]pyridine

To a solution of the product from Step A (480 mg, 2.18 mmol) in ethanol (12 mL) was added chloroacetaldehyde (50 wt.% in water, 0.336 mL) and the mixture was heated at 85°C for 3 hours. After cooling to room temperature, the solution was concentrated under reduced pressure. The residue was partitioned between saturated aqueous sodium bicarbonate and dichloromethane. The organic phase was separated, dried (magnesium sulfate) and concentrated in vacuo to give the product as a light brown solid which was used without further purification. LC/MS 244.8 (M+1).

Intermediate 7

5-(3-Iodophenyl)-1,3,4-oxadiazol-2(3H)-one

Step A: 3-Iodophenhydrazide

A suspension of 3-iodobenzoic acid (1.24 g, 5.0 mmol) in 15 mL of dichloromethane containing 1 drop of N,N-dimethylformamide was treated dropwise with oxalyl chloride (0.70 mL, 7.5 mmol). After stirring at room temperature for 2.5 hours, the solution was concentrated under reduced pressure to obtain a light orange oil, which was dissolved in 10 mL of anhydrous tetrahydrofuran (THF), and added dropwise to tert-butyl carbazate (793 mg, 6.0 mmol) and tris Ethylamine (1.10 mL) was ice-cooled suspension in 15 mL dry THF. The resulting mixture was allowed to warm to room temperature overnight, then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and water, and the organic layer was washed sequentially with 5% aqueous hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure to give an off-white solid. Trituration with hexanes gave a white powder. It was suspended in 60 mL of dichloromethane and cooled in an ice-water bath. Trifluoroacetic acid (25 mL) was added dropwise and the mixture was allowed to warm to room temperature. After 1 hour, the solution was concentrated under reduced pressure, the residue was dissolved in 60 mL of water, and neutralized to pH 8 with 1N aqueous sodium hydroxide solution. The resulting precipitate was collected, washed with water, and dried in vacuo to give 3-iodophenylhydrazide as a white powder.

Step B: 5-(3-Iodophenyl)-1,3,4-oxadiazol-2(3H)-one

A solution of N,N-carbonyldiimidazole (800 mg, 5.0 mmol) in 5 mL of anhydrous THF was added dropwise to the product from Step A (1.048 g, 4.0 mmol) and triethylamine (0.60 mL, 4.0 mmol) in 12 mL of Ice-cooled solution in water THF. The resulting mixture was allowed to warm to room temperature overnight, then concentrated under reduced pressure. The residue was partitioned between ether and water, and the ether layer was washed sequentially with 5% aqueous hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure to give a white powder. Recrystallization from ethyl acetate-hexanes gave the title compound as a fluffy white solid. LC/MS 289.2 (M+1).

Intermediate 8

5-ethoxy-3-(3-iodophenyl)-1H-1,2,4-triazole

Step A: 2-Amino-5-(3-iodophenyl)-1,2,4-oxadiazole

A solution of cyanogen bromide (1.06 g, 10.0 mmol) in 10 mL of methanol was added dropwise to an ice-cooled suspension of 3-iodophenylhydrazide (2.62 g, 10.0 mmol, Intermediate 7, Step A) in 20 mL of methanol . After 30 minutes, the mixture was allowed to warm to room temperature, then warmed at reflux for 1.5 hours. The resulting solution was cooled to 0°C and neutralized to pH 9 with concentrated aqueous ammonium hydroxide. The resulting precipitate was collected, washed with methanol, and dried in vacuo to give the product as an off-white powder. LC/MS 288.0 (M+1).

Step B: 5-Ethoxy-3-(3-iodophenyl)-1H-1,2,4-triazole

The product of Step A (1.00 g, 3.48 mmol) was added to a solution of potassium hydroxide (1.0 g) in 30 mL of absolute ethanol. The mixture was warmed to reflux and after 5 hours cooled to room temperature. The solution was then acidified with glacial acetic acid and concentrated under reduced pressure. The product was extracted with ethyl acetate and the combined ethyl acetate extracts were washed with water followed by saturated brine, dried (magnesium sulfate) and concentrated to give the crude product as an orange semi-solid. Purification by flash chromatography (silica gel, 15% ethyl acetate-hexanes) gave the product as an off-white sticky foam. LC/MS 316.0 (M+1).

Intermediate 9

4-(3-Bromophenyl)-1,3-dihydro-2H-imidazol-2-one

To a mixture of 3-bromophenacyl bromide (0.5 g, 1.8 mmol), urea (0.32 g, 5.3 mmol) and ammonium acetate (0.40 g, 5.4 mmol) in water (10 mL) was added glacial acetic acid (0.32 g, 5.4 mmol). The reaction mixture was heated at reflux overnight. After cooling to room temperature, the mixture was extracted with three portions of ethyl acetate, the combined extracts were dried (magnesium sulfate) and concentrated under reduced pressure to give a brown residue which was triturated with ether to give the desired product. LC/MS 238.9 and 240.9 (M+1).

Intermediate 10

3-(3-iodophenyl)-5-(trifluoromethyl)-4H-1,2,4-triazole

Step A: Ethyl 3-iodobenzimidate hydrochloride

Hydrogen chloride gas was bubbled through a solution of 3-iodobenzonitrile (2.00 g, 8.73 mmol) in 20 mL of absolute ethanol for 30 minutes at room temperature. The resulting solution was kept at room temperature for 48 hours, then concentrated under reduced pressure. The residue was triturated with ether, collected and dried in vacuo to give the product as a white powder.

Step B: 3-iodobenzamidine hydrochloride

Ammonia gas was bubbled through an ice-cooled suspension of the product from Step A (1.00 g, 3.21 mmol) in 20 mL of absolute ethanol for 20 minutes. The resulting clear solution was allowed to warm to room temperature and stir for 48 hours. The reaction mixture was then concentrated under reduced pressure and the residue was triturated with ether. The supernatant was decanted and the residual gummy product was dried in vacuo to give the title compound as a white foam.

Step C: 3-(3-Iodophenyl)-5-(trifluoromethyl)-4H-1,2,4-triazole

To a solution of ethyl trifluoroacetate (0.12 mL, 1.00 mmol) in 4.0 mL of anhydrous tetrahydrofuran was added anhydrous hydrazine (25 mL, 0.80 mmol), and the resulting solution was heated to reflux. After 1 hour, the solution was cooled to room temperature and added via syringe to the mixture of amidine hydrochloride (283 mg, 1.00 mmol) from Step B and solid sodium hydroxide (50 mg) in 3.0 mL of anhydrous THF. The reaction mixture was warmed at reflux for 3 hours, then cooled to room temperature overnight. The precipitated solid was removed by filtration and the filtrate was concentrated under reduced pressure to give a yellow gum which was partitioned between ethyl acetate and water. The organic layer was washed with saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (silica gel, step gradient, 0-20% ethyl acetate/hexanes) to give a white solid product of. LC/MS 339.8 (M+1).

Intermediate 11

6-iodoquinazolin-4(3H)-one

A mixture of 2-amino-5-iodobenzoic acid (2.0 g, 7.6 mmol) and formamidine acetate (0.99 g, 9.5 mmol) in absolute ethanol (80 mL) was heated at reflux for 2 hours. The reaction was then cooled in an ice bath and water (10 mL) was added with stirring. The resulting precipitate was collected by filtration to give the desired product as pale crystals. LC/MS 272.7 (M+1).

Intermediate 12

6-iodo-3-(trifluoromethyl)[1,2,4]triazolo[4,3-α]pyridine

Step A: 2,2,2-Trifluoro-N'-(5-iodopyridin-2-yl)acetylhydrazide

To a solution of 5-iodo-2-hydrazinopyridine (235 mg, 1 mmol) in trifluoroacetic acid (1.5 mL) in a thick wall sealable tube was added trifluoroacetic anhydride (0.356 mL, 2.5 mmol). The mixture was warmed to 50°C. After 18 hours, the mixture was concentrated to dryness under reduced pressure. The resulting residue was purified by preparative thin layer chromatography (silica gel, 5% methanol/dichloromethane) to afford the product. LC/MS 331.8 (M+1).

Step B: 6-iodo-3-(trifluoromethyl)[1,2,4]triazolo[4,3-α]pyridine

The product of Step A (145mg, 0.438mmol) was suspended in superphosphoric acid (5mL) and the mixture was heated at 140°C for 6 hours. The mixture was cooled to room temperature, poured into ice, and neutralized with concentrated aqueous ammonium hydroxide. The mixture was extracted several times with ethyl acetate, and the combined extracts were washed with saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure. The resulting residue was purified by preparative thin layer chromatography (silica gel, 8% methanol/dichloromethane) to afford the title compound. LC/MS 313.8 (M+1).

Intermediate 13

6-iodo[1,2,4]triazolo[4,3-α]pyridin-3(2H)-one

To a solution of 5-iodo-2-hydrazinopyridine (470 mg, 2 mmol) in dichloromethane (30 mL) was added a solution of N,N-carbonyldiimidazole (389 mg, 2.4 mmol) in 20 mL of dichloromethane. After stirring at room temperature for 2 hours, the mixture was washed successively with water, 0.5N aqueous hydrochloric acid solution and saturated aqueous sodium bicarbonate solution. The organic phase was dried (magnesium sulfate) and concentrated under reduced pressure to give the product. The aqueous phase was left to settle overnight, and the resulting yellow crystals were collected and dried in vacuo to obtain the product. LC/MS 261.8 (M+1).

Intermediate 14

6-iodo-2-methyl[1,2,4]triazolo[4,3-α]pyridin-3(2H)-one

To 6-iodo[1,2,4]triazolo[4,3-α]pyridin-3(2H)-one (Intermediate 13, 360mg, 1.379mmol) in anhydrous dimethylformamide (DMF, To a solution in 4 mL) was added successively cesium carbonate (1.35 g, 4.14 mmol) and iodomethane (0.52 mL), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was then diluted with ethyl acetate and the precipitated solid was removed by filtration. The filtrate was washed with saturated brine, dried (magnesium sulfate) and concentrated to dryness under reduced pressure. The residue was purified by preparative thin layer chromatography (silica gel, 3% methanol/dichloromethane) to afford the title compound.

LC/MS 257.8 (M+1).

Intermediate 15

7-iodoquinazolin-4(3H)-one

Step A: 4-Chloro-6-iodoquinazoline

A mixture of 6-iodoquinazolin-4(3H)-one (Intermediate 11, 100 mg, 0.37 mmol) and phenylphosphonodichloride (1.5 mL) was heated at 150 °C for 1 hour. After cooling the reaction mixture in an ice bath, isopropyl ether was added. The resulting crystalline precipitate was collected by filtration, and then stirred with saturated aqueous sodium bicarbonate solution. The solution was extracted with three portions of ethyl acetate and the combined extracts were dried (magnesium sulfate) and concentrated under reduced pressure to give the crude product. Purification by preparative thin layer chromatography (silica gel, 5% and 10% ethyl acetate/hexanes) afforded the desired product. LC/MS 291 (M+1).

Step B: 7-iodoquinazolin-4(3H)-one

To the product of Step A (73.8 mg) was added 2M ammonia in ethanol (7 mL) and the resulting solution was stirred at room temperature for 2 hours. The volatiles were removed under reduced pressure and the product was triturated with ether to give the desired product as a white powder. LC/MS 271.8 (M+1).

Intermediate 16

N-(6-iodoimidazo[1,2-α]pyridin-2-yl)acetamide

Step A: 5-iodo-2-p-toluenesulfonylaminopyridine

p-Toluenesulfonyl chloride (2.00 g, 10.4 mmol) was added to a solution of 5-iodo-2-aminopyridine (2.50 g, 11.4 mmol) in 6 mL of pyridine, and the resulting solution was warmed at 90° C. for 18 hours. After cooling to room temperature, the solution was added portionwise to 75 mL of ice water with stirring. The resulting precipitate was collected, washed with water, and dried in vacuo to give a pale yellow powder, which was stirred with 20 mL of methanol for several minutes. The solid product was then collected, washed with methanol, and dried in vacuo to afford the title compound as an off-white powder. LC/MS 374.8 (M+1).

Step B: 1-(carbamoylmethyl)-5-iodo-2-(p-toluenesulfonylamino)pyridine

To a solution of the product from Step A (1.60 g, 4.27 mmol) in 8.0 mL of anhydrous N,N-dimethylformamide was added sodium hydride (188 mg, 60% by weight in mineral oil, 4.44 mmol) . After 15 minutes, the solution was warmed to 60°C for 10 minutes and then cooled to room temperature. Chloroacetamide (420mg) was added in one portion, then the solution was warmed to 100°C. After 2.5 hours, the solution was cooled to room temperature and poured into 70 mL of ice water. The resulting precipitate was collected, washed with water and air dried overnight. The crude product was then stirred with 20 mL of methanol for several minutes and the product was collected by filtration to give the title compound as a white powder. LC/MS 414.9 (M+1- _H2O ).

Step C: N-(6-iodoimidazo[1,2-α]pyridin-2-yl)acetamide

A mixture of the product of Step B (300 mg, 0.70 mmol) and 1.0 mL of acetic anhydride was warmed at reflux for 2.5 hours, the solution was cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between ethyl acetate and water, and the ethyl acetate layer was washed successively with saturated aqueous sodium bicarbonate and saturated brine, dried (magnesium sulfate) and concentrated under reduced pressure. The crude residue was purified by preparative thin layer chromatography (silica gel, 7% methanol/dichloromethane) to afford the title compound as a light yellow powder. LC/MS 301.9 (M+1).

Intermediate 17

6-iodo-3-nitroimidazo[1,2-α]pyridine

To a solution of 6-iodoimidazo[1,2-α]pyridine (Intermediate 6, 448 mg, 1.84 mmol) in concentrated sulfuric acid (1.8 mL) was added concentrated nitric acid (0.54 mL) dropwise at 15°C. After the addition was complete, the reaction mixture was stirred at room temperature for 1 hour, then poured onto 10 g of ice. The pH of the mixture was adjusted to 4 with aqueous potassium hydroxide and the resulting solid was collected by filtration, washed with water and dried. The crude product was recrystallized from dichloromethane/hexanes to afford the title compound. LC/MS 289.8 (M+1).

Intermediate 18

6-Bromo[1,2,4]triazolo[1,5-α]pyridine

Step A: N'-(5-bromopyridin-2-yl)-N,N-dimethyliminoformamide

To a stirred solution of 5-bromo-2-aminopyridine (3.0 g, 17.3 mmol) in N,N-dimethylformamide (6 mL) was added N,N-dimethylformamide dimethyl acetal (5.37 g, 45.0 mmol). The reaction mixture was heated to 130°C overnight. After cooling to room temperature, the volatiles were removed under reduced pressure to give the desired product as a brown oil. LC/MS 227.8 (M+1).

Step B: 6-Bromo[1,2,4]triazolo[1,5-α]pyridine

To an ice-cooled stirred solution of the crude product from Step A (3.94 g, 17.3 mmol) in methanol (30 mL) and pyridine (2.73 g, 35.6 mmol) was added hydroxylamine-O-sulfonic acid (2.54 g, 22.5 mmol). The reaction mixture was allowed to warm to room temperature and stirred overnight. The volatiles were removed under reduced pressure and the residue was partitioned between aqueous sodium bicarbonate and ethyl acetate. The aqueous layer was further extracted with ethyl acetate and the combined organic layers were washed successively with water (100 mL) and saturated brine solution (100 mL), dried (magnesium sulfate) and concentrated in vacuo to give a brown solid which was recrystallized from dichloromethane to give The title compound as an orange solid. LC/MS 197.9 and 199.9 (M+1).

Intermediate 19

7-iodo[1,2,4]triazolo[1,5-α]pyridine

Step A: 2-(tert-Butoxycarbonyl)amino-4-iodopyridine

To 4-iodopicolinic acid hemi-hydroiodide hydrate (Lohse, O.Synth.Commun.1996, 26, 2017; 24.5g, 78.3mmol) in 140mL tert-butanol, 130mL toluene and 35mL triethyl To the stirred solution in the amine was added diphenylphosphoryl azide (27 mL, 125 mmol) dropwise over 30 minutes. The resulting solution was then warmed to 65°C and after 1.5 hours the bath temperature was raised to 100°C. After 4 hours, the solution was cooled and concentrated under reduced pressure. The residue was partitioned between ethyl acetate (600 mL) and water (300 mL). The ethyl acetate layer was washed sequentially with saturated aqueous sodium bicarbonate (200 mL) and saturated brine (200 mL), dried over magnesium sulfate and concentrated to give a brown solid. Purification by flash chromatography (silica gel, 5% ethyl acetate-hexane eluent) gave a pale yellow solid which was triturated with hexane to give the title compound as a white solid. MS 265.1 (M+1-tert-butyl).

Step B: 2-Amino-4-iodopyridine

To an ice-cooled solution of the product from Step A above (3.84 g, 12.0 mmol) in 25 mL of dichloromethane was added trifluoroacetic acid (12 mL) dropwise. The resulting solution was allowed to warm to room temperature and after 1 h the volatiles were removed under reduced pressure. The residue was dissolved in water (120 mL), and sodium bicarbonate was added portionwise to neutralize the solution. The mixture was extracted with ethyl acetate, the extract was washed with saturated brine, dried over magnesium sulfate and concentrated to give an off-white solid which was triturated with hexanes to give the title compound as a white powder. MS 221.1(M+1).

Step C: 7-iodo[1,2,4]triazolo[1,5-α]pyridine

To a stirred solution of the product from Step B above (220 mg, 1.00 mmol) in N,N-dimethylformamide (0.5 mL) was added N,N-dimethylformamide dimethyl acetal (0.37 mL, 2.60 mmol). The reaction mixture was heated to 130°C overnight. After cooling to room temperature, the volatiles were removed under reduced pressure to give a red oil which was dissolved in 2.0 mL of methanol and 0.162 mL of pyridine. The solution was cooled in an ice bath and hydroxylamine-O-sulfonic acid (147 mg, 1.30 mmol) was added in one portion. The reaction mixture was allowed to warm to room temperature and stirred overnight. The volatiles were removed under reduced pressure and the residue was partitioned between saturated brine solution and ethyl acetate. The aqueous layer was further extracted with ethyl acetate, the combined organic layers were washed with saturated brine solution (100 mL), dried over magnesium sulfate and concentrated under reduced pressure to give the title compound as an orange solid. MS 246.1(M+1).

Intermediate 20

5-Chloropyrazolo[1,5-α]pyrimidine

Step A: 5-Hydroxypyrazolo[1,5-α]pyrimidine

To a stirred solution of 3-aminopyrazole (11.5 g, 0.138 mmol) in 65 mL of 1,4-dioxane was added dropwise ethyl propiolate (14.7 g, 15.2 mL, 0.150 mol), and the resulting light yellow The solution was warmed to reflux. After 4 hours, the blood-red solution was cooled to room temperature, and 100 mL of toluene was added with stirring. The resulting precipitate was collected, washed with toluene and air dried to give the title compound as a tan solid. MS 136.0(M+1).

Step B: 5-Chloropyrazolo[1,5-α]pyrimidine

A mixture of the product from Step A above (1.35 g, 10.0 mmol) and 7.5 mL of phosphorus oxychloride was warmed at reflux for 4 hours. The mixture was cooled and volatiles were removed under reduced pressure. The dark residue was partitioned between ice water and dichloromethane, and the aqueous layer was extracted with additional dichloromethane. The combined organic extracts were dried over sodium sulfate and concentrated under reduced pressure to give a light brown solid which was purified by flash chromatography (silica gel; 0.5% methanol in dichloromethane eluent) to give the title compound as a white solid. MS 153.8 and 155.8 (M+1).

The Examples listed in Tables 1-3 were prepared essentially according to the procedures outlined in Examples 1-17.

Table 1

Example R ⁴ R ² x MS(M+1) 18 4-(SO ₂ Me)-phenyl Me (S)-CHF 405.1 19 3-(SO ₂ Me)-phenyl Me (S)-CHF 405.1 20 Pyrazin-5-yl Me (S)-CHF 329.2 twenty one 3-chloropyridin-4-yl Me (S)-CHF 362.1 twenty two 2,4-Difluorophenyl Me (S)-CHF 363.0 twenty three 3,4-Difluorophenyl Me (S)-CHF 363.1 twenty four 2,5-Difluorophenyl Me (S)-CHF 363.0 25 3,5-difluorophenyl Me (S)-CHF 363.0 26 3-(ethoxycarbonyl)phenyl Me (S)-CHF 399.2 27 4-(ethoxycarbonyl)phenyl Me (S)-CHF 399.1 28 3-(NHSO ₂ Me)-phenyl Me (S)-CHF 420.1 29 4-(NHSO ₂ Me)-phenyl Me (S)-CHF 420.1 30 4-CO ₂ H-phenyl Me (S)-CHF 371.0 31 Pyridin-3-yl Me (S)-CHF 328.1 32 6-OMe-pyridin-3-yl Me (S)-CHF 358.1 33 2-Cl-phenyl Me (S)-CHF 361.1 34 2-F-phenyl Me (S)-CHF 345.1 35 3-CN-phenyl Me (S)-CHF 352.1 36 Pyridin-4-yl Me (S)-CHF 328.1 37 Pyridin-2-yl Me (S)-CHF 328.1 38 2,4-Difluorophenyl Cyclopropylmethyl (S)-CHF 403.1 39 3-(MeSO ₂ )-phenyl Cyclopropylmethyl (S)-CHF 444.9 40 4-fluoro-(3-tetrazol-5-yl)phenyl Me (S)-CHF 413.1 41 3-(aminosulfonyl)phenyl Me (S)-CHF 406.0 42 2-Fluoro-(5-tetrazol-5-yl)phenyl Me (S)-CHF 413.3 43 3-Hydroxyphenyl Me (S)-CHF 343.1 44 6-fluoropyridin-3-yl Me (S)-CHF 346.2 45 3-(Aminocarbonyl)phenyl Me (S)-CHF 370.2 46 3-(Phenylaminocarbonyl)phenyl Me (S)-CHF 446.3 47 4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl Me (S)-CHF 429.2 48 3-[(thiazol-2-yl)aminocarbonyl]phenyl Me (S)-CHF 453.2 49 3-[(tetrazol-5-yl)aminocarbonyl]phenyl Me (S)-CHF 438.2 50 Imidazo[1,2-a]pyridin-6-yl Me (S)-CHF 367.2 51 2-Methoxyphenyl Me (S)-CHF 357.2 52 3-(5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)phenyl Me (S)-CHF 411.1 53 3-(5-ethoxy-1H-1,2,4-triazol-3-yl)phenyl Me (S)-CHF 438.3 54 1-Ethyl-6-oxo-1,6-dihydropyridin-3-yl Me (S)-CHF 372.2 55 Quinolin-6-yl Me (S)-CHF 378.2 56 3-(2-oxo-2,3-dihydro-1H-imidazol-4-yl)phenyl Me (S)-CHF 409.3 57 2-Methylphenyl Me (S)-CHF 341.2 58 2-(trifluoromethyl)phenyl Me (S)-CHF 395.1 59 3-[5-(trifluoromethyl)-4H-1,2,4-triazol-3-yl]phenyl Me (S)-CHF 462.1 60 4-oxo-3,4-dihydroquinazolin-6-yl Me (S)-CHF 395.1 61 4-fluorophenyl CONHEt (S)-CHF 402.2 62 1-methyl-6-oxo-1,6-dihydropyridin-3-yl Et (S)-CHF 372.1 63 6-oxo-1,6-dihydropyridin-3-yl Et (S)-CHF 358.1 64 3-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-6-yl Me (S)-CHF 436.2 65 4-fluorophenyl CONH ₂ (S)-CHF 374.2 66 3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-a]pyridin-6-yl Me (S)-CHF 384.1 67 1-methyl-6-oxo-1,6-dihydropyridin-3-yl Me CF ₂ 376.0 68 6-oxo-1,6-dihydropyridin-3-yl Me CF ₂ 362.0 69 4-Aminoquinazolin-6-yl Me (S)-CHF 394.1 70 5-Bromo-1-methyl-6-oxo-1,6-dihydropyridin-3-yl Me (S)-CHF 423.9 71 4-fluorophenyl (Pyrrolidin-1-yl)carbonyl (S)-CHF 428.3 72 4-fluorophenyl (Azetidin-1-yl)carbonyl (S)-CHF 414.3 73 2-(acetylamino)imidazo[1,2-a]pyridin-6-yl Me (S)-CHF 424.2 74 2-Methyl-3-oxo-2,3-dihydro[1,2,4]triazolo[4,3-a]pyridin-6-yl Me (S)-CHF 398.1 75 3-Aminoimidazo[1,2-a]pyridin-6-yl Me (S)-CHF 382.1 76 4-fluorophenyl [(tetrazol-5-yl)amino]carbonyl (S)-CHF 442.3 77 4-fluorophenyl CONHMe (S)-CHF 388.2 78 4-fluorophenyl CONEt ₂ (S)-CHF 430.3 79 4-fluorophenyl COOMe (S)-CHF 389.2 80 [1,2,4]triazolo[1,5-a]pyridin-6-yl Me (S)-CHF 368.1 81 4-fluorophenyl COOH CH ₂ 357.1 82 4-fluorophenyl COOH CF ₂ 393.0 83 4-fluorophenyl CONMe ₂ CH ₂ 384.1 84 3-carboxypyrazolo[1,5-a]pyridin-5-yl Me (S)-CHF 411.1 85 6-oxo-1,6-dihydropyridin-3-yl Me CH ₂ 326.1 86 [1,2,4]triazolo[1,5-a]pyridin-6-yl CONMe ₂ (S)-CHF 425.3 87 [1,2,4]triazolo[1,5-a]pyridin-7-yl CONMe ₂ (S)-CHF 425.3 88 Pyrazolo[1,5-a]pyrimidin-5-yl CONMe ₂ (S)-CHF 425.2 89 Phenyl Me (S)-CHF 327.0 90 [1,2,4]triazolo[1,5-a]pyridin-6-yl CONMe ₂ CH ₂ 407.4 91 [1,2,4]triazolo[1,5-a]pyridin-6-yl (3-Fluoroazetidin-1-yl)carbonyl CH ₂ 441.2 92 [1,2,4]triazolo[1,5-a]pyridin-6-yl (Pyrrolidin-1-yl)carbonyl CF ₂ 469.4 93 [1,2,4]triazolo[1,5-a]pyridin-6-yl CONMeCH ₂ -Ph CF ₂ 519.1 94 [1,2,4]triazolo[1,5-a]pyridin-6-yl (morpholin-4-yl)carbonyl CF ₂ 485.2

Table 2

Example R ⁴ R ² x MS(M+1) 95 4-fluorophenyl Me CHF 331.2 96 1-methyl-6-oxo-1,6-dihydropyridin-3-yl Me CHF 344.0 97 [1,2,4]triazolo[4,3-a]pyridin-6-yl Me CHF 354.2 98 Imidazo[1,2-a]pyridin-6-yl Me CHF 353.1 99 4-fluorophenyl COOH CHF 361.1 100 4-fluorophenyl CONMe ₂ CHF 388.1 101 [1,2,4]triazolo[1,5-a]pyridin-6-yl CONMe ₂ CHF 443.2

table 3

Example R ⁴ R ² x MS(M+1) 102 4-fluorophenyl Me CHF 359.2 103 1-methyl-6-oxo-1,6-dihydropyridin-3-yl Me CHF 372.1 104 [1,2,4]triazolo[4,3-a]pyridin-6-yl Me CHF 382.1 105 Imidazo[1,2-a]pyridin-6-yl Me CHF 381.1

Examples of pharmaceutical preparations

As a specific example of an oral pharmaceutical composition, a 100 mg effective tablet consists of 100 mg of any compound of the invention, 268 mg of microcrystalline cellulose, 20 mg of croscarmellose sodium and 4 mg of magnesium stearate. The active ingredient, microcrystalline cellulose and croscarmellose are first mixed. The mixture is then lubricated with magnesium stearate and compressed into tablets.

While the present invention has been described and illustrated with reference to certain specific embodiments thereof, those skilled in the art will appreciate that various modifications, changes, modifications, substitutions, deletions or additions may be made in various methods and schemes without departing from the present invention spirit and scope. For example, effective dosages other than those specifically described herein above may be used as a result of variations in the response of mammals to treatment with the compounds of the invention described above for any of their indications. According to and based on the specific active compound selected or the presence or absence of a pharmaceutical carrier, as well as the type of formulation and the mode of administration used, the specific pharmacological response observed may vary, and such expected changes or differences in results are included in the present invention. purpose and practice. Accordingly, the invention should be defined only by the scope of the following claims, and the claims should be interpreted as broadly as reasonably possible.

Claims (34)

1. a structural formula I compound or its pharmacy acceptable salt:

Wherein

Each n independently is 0,1 or 2;

M and p independently are 0 or 1 separately;

X is CH ₂, S, CHF or CF ₂

W and Z independently are CH separately ₂, CHF or CF ₂

R ¹Be hydrogen or cyano group;

Each R ³Independently be selected from hydrogen, halogen, C _1-4Alkyl, C _1-4Alkoxyl group, cyano group, trifluoromethyl, trifluoromethoxy and hydroxyl;

R ⁴Be aryl, heteroaryl or heterocyclic radical, wherein aryl, heteroaryl and heterocyclic radical do not replace or with 1-5 R ⁵Substituting group replaces;

R ²Be selected from

Hydrogen,

C _1-10Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen or hydroxyl,

C _2-10Thiazolinyl, wherein thiazolinyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen or hydroxyl,

(CH ₂) _n-aryl, wherein aryl does not replace or independently is selected from hydroxyl, halogen, CO with 1-5 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-heteroaryl, wherein heteroaryl does not replace or independently is selected from hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-heterocyclic radical, wherein heterocyclic radical does not replace or independently is selected from oxo, hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-C _3-6Cycloalkyl, wherein cycloalkyl does not replace or independently is selected from halogen, hydroxyl, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _nCOOH，

(CH ₂) _nCOOC _1-6Alkyl,

(CH ₂) _nCONR ⁶R ⁷, R wherein ⁶And R ⁷Independently be selected from hydrogen, tetrazyl, thiazolyl, (CH ₂) _n-phenyl, (CH ₂) _n-C _3-6Cycloalkyl and C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen and hydroxyl, and phenyl and cycloalkyl does not replace or independently be selected from halogen, hydroxyl, C with 1-5 wherein _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen;

Perhaps R wherein ⁶And R ⁷The nitrogen-atoms that connects with them forms the heterocycle that is selected from azetidine, tetramethyleneimine, piperidines, piperazine and morpholine, and wherein said heterocycle does not replace or independently is selected from halogen, hydroxyl, C with 1-3 _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen;

R wherein ²In any methylene radical (CH ₂) carbon atom do not replace or independently be selected from halogen, hydroxyl and do not replace or the C of 1-5 halogen replacement with 1-2 _1-4The group of alkyl replaces;

Each R ⁵Independently be selected from

Halogen,

Cyano group,

Oxo,

Hydroxyl,

C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 halogen,

C _1-6Alkoxyl group, wherein alkoxyl group does not replace or replaces with 1-5 halogen,

(CH ₂) _n-NR ⁶R ⁷，

(CH ₂) _n-CONR ⁶R ⁷，

(CH ₂) _n-OCONR ⁶R ⁷，

(CH ₂) _n-SO ₂NR ⁶R ⁷，

(CH ₂) _n-SO ₂R ⁹，

(CH ₂) _n-NR ⁸SO ₂R ⁹，

(CH ₂) _n-NR ⁸CONR ⁶R ⁷，

(CH ₂) _n-NR ⁸COR ⁸，

(CH ₂) _n-NR ⁸CO ₂R ⁹，

(CH ₂) _n-COOH，

(CH ₂) _n-COOC _1-6Alkyl,

(CH ₂) _n-aryl, wherein aryl does not replace or independently is selected from halogen, hydroxyl, CO with 1-5 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl, C _3-6Cycloalkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-heteroaryl, wherein heteroaryl does not replace or independently is selected from hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl, C _3-6Cycloalkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-heterocyclic radical, wherein heterocyclic radical does not replace or independently is selected from oxo, hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl, C _3-6Cycloalkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-C _3-6Cycloalkyl, wherein cycloalkyl does not replace or independently is selected from halogen, hydroxyl, C with 1-3 _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

R wherein ⁵In any methylene radical (CH ₂) carbon atom do not replace or independently be selected from halogen, hydroxyl and do not replace or with the C of 1-5 halogen replacement with 1-2 _1-4The group of alkyl replaces;

Each R ⁹Independently be selected from tetrazyl, thiazolyl, (CH ₂) _n-phenyl, (CH ₂) _n-C _3-6Cycloalkyl and C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 halogen, and phenyl and cycloalkyl does not replace or independently be selected from halogen, hydroxyl, C with 1-5 wherein _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or with 1-5 halogen replacement, and R wherein ⁸In any methylene radical (CH ₂) carbon atom do not replace or independently be selected from halogen, hydroxyl and do not replace or with the C of 1-5 halogen replacement with 1-2 _1-4The group of alkyl replaces;

Each R ⁸Be hydrogen or R ⁹

2. the compound of claim 1 wherein has the three-dimensional chemical configuration described in the formula Ia with the * marked carbon atoms:

R ³Be hydrogen or fluorine.

3. the compound of claim 2 is wherein with the R that is connected in of * * mark ¹Carbon atom have the three-dimensional chemical configuration described in the formula Ib:

4. the compound of the structural formula Ic of claim 1:

R wherein ³Be hydrogen or fluorine.

5. the compound of claim 4 wherein has the three-dimensional chemical configuration described in the structural formula Id with the * marked carbon atoms:

6. the compound of claim 5, wherein R ¹Be hydrogen, W is CH ₂, X is CH ₂, CHF or CF ₂

7. the compound of the structural formula Ie of claim 1:

R wherein ³Be hydrogen or fluorine.

8. the compound of claim 7 wherein has the three-dimensional chemical configuration described in the structural formula If with the * marked carbon atoms:

9. the compound of the structural formula Ig of claim 1:

R wherein ³Be hydrogen or fluorine.

10. the compound of claim 9 wherein has the three-dimensional chemical configuration described in the structural formula Ih with the * marked carbon atoms:

11. the compound of claim 10, wherein W and Z are CH ₂, X is CH ₂, CHF or CF ₂

12. the compound of claim 1, wherein R ²Be selected from

C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen or hydroxyl,

C _2-6Thiazolinyl, wherein thiazolinyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen or hydroxyl,

(CH ₂) _n-C _3-6Cycloalkyl, wherein cycloalkyl does not replace or independently is selected from halogen, hydroxyl, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _nCOOH，

(CH ₂) _nCOOC _1-6Alkyl,

(CH ₂) _nCONR ⁶R ⁷, R wherein ⁶And R ⁷Independently be selected from hydrogen, tetrazyl, thiazolyl, (CH ₂) _n-phenyl, (CH ₂) _n-C _3-6Cycloalkyl and C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen and hydroxyl, and phenyl and cycloalkyl does not replace or independently be selected from halogen, hydroxyl, C with 1-5 wherein _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen;

Perhaps R wherein ⁶And R ⁷The nitrogen-atoms that connects with them forms the heterocycle that is selected from azetidine, tetramethyleneimine, piperidines, piperazine and morpholine, and wherein said heterocycle does not replace or independently is selected from halogen, hydroxyl, C with 1-5 _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen;

R wherein ²In any methylene radical (CH ₂) carbon atom do not replace or independently be selected from halogen, hydroxyl and do not replace or with the C of 1-5 halogen replacement with 1-2 _1-4The group of alkyl replaces.

13. the compound of claim 12, wherein R ²Be selected from

C _1-3Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen or hydroxyl,

CH ₂-C _3-6Cycloalkyl,

COOH，

COOC _1-6Alkyl,

CONR ⁶R ⁷, R wherein ⁶And R ⁷Independently be selected from hydrogen, tetrazyl, thiazolyl, (CH ₂) _n-phenyl, (CH ₂) _n-C _3-6Cycloalkyl and C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 substituting group that independently is selected from halogen and hydroxyl, and phenyl and cycloalkyl does not replace or independently be selected from halogen, hydroxyl, C with 1-5 wherein _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen;

Perhaps R wherein ⁶And R ⁷The nitrogen-atoms that connects with them forms the heterocycle that is selected from azetidine, tetramethyleneimine, piperidines, piperazine and morpholine, and wherein said heterocycle does not replace or independently is selected from halogen, hydroxyl, C with 1-5 _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen.

14. the compound of the structural formula Ii of claim 1:

Wherein X is CH ₂, S, CHF or CF ₂

W and Z independently are CH separately ₂, CHF or CF ₂

R ⁴Be phenyl, heteroaryl or heterocyclic radical, wherein phenyl, heteroaryl and heterocyclic radical do not replace or with 1-3 R ⁵Substituting group replaces;

R ²Be selected from:

Methyl,

Ethyl,

CH ₂-cyclopropyl,

COOH，

COOMe，

COOEt，

CONHMe，

CONMe ₂，

CONH ₂，

CONHEt，

CONMeCH ₂Ph，

Tetramethyleneimine-1-base carbonyl,

Azetidine-1-base carbonyl,

3-fluorine azetidine-1-base carbonyl,

Morpholine-4-base carbonyl,

[(tetrazolium-5-yl) amino] carbonyl;

Each R ⁵Independently be selected from:

Halogen,

Cyano group,

Oxo,

Hydroxyl,

C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 halogen,

C _1-6Alkoxyl group, wherein alkoxyl group does not replace or replaces with 1-5 halogen,

NR ⁶R ⁷，

CONR ⁶R ⁷，

OCONR ⁶R ⁷，

SO ₂NR ⁶R ⁷，

SO ₂R ⁹，

NR ⁸SO ₂R ⁹，

NR ⁸CONR ⁶R ⁷，

NR ⁸COR ⁸，

NR ⁸CO ₂R ⁹，

COOH，

COOC _1-6Alkyl,

Aryl, wherein aryl does not replace or independently is selected from halogen, hydroxyl, CO with 1-5 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

Heteroaryl, wherein heteroaryl does not replace or independently is selected from hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

Heterocyclic radical, wherein heterocyclic radical does not replace or independently is selected from oxo, hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

(CH ₂) _n-C _3-6Cycloalkyl, wherein cycloalkyl does not replace or independently is selected from halogen, hydroxyl, C with 1-3 _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen.

15. the compound of claim 14, wherein each R ⁵Independently be selected from:

Halogen,

Cyano group,

Oxo,

C _1-6Alkyl, wherein alkyl does not replace or replaces with 1-5 halogen,

C _1-6Alkoxyl group, wherein alkoxyl group does not replace or replaces with 1-5 halogen,

CONR ⁶R ⁷，

NR ⁸COR ⁸，

SO ₂R ⁹，

NR ⁸SO ₂R ⁹，

COOH，

COOC _1-6Alkyl,

Heteroaryl, wherein heteroaryl does not replace or independently is selected from hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen,

Heterocyclic radical, wherein heterocyclic radical does not replace or independently is selected from oxo, hydroxyl, halogen, CO with 1-3 ₂H, C _1-6Alkoxy carbonyl, C _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces, and wherein alkyl and alkoxyl group do not replace or replace with 1-5 halogen.

16. the compound of claim 14, wherein R ⁴Be selected from:

The 4-fluorophenyl,

The 2,4 difluorobenzene base,

3, the 4-difluorophenyl,

The 2-chloro-phenyl-,

The 2-fluorophenyl,

3-(methyl sulphonyl) phenyl,

3-(ethoxy carbonyl) phenyl,

The 3-carboxyl phenyl,

3-(aminocarboxyl) phenyl,

3-[(tertiary butyl amino) carbonyl] phenyl,

The 3-[(phenyl amino) carbonyl] phenyl,

3-[(thiazol-2-yl amino) carbonyl] phenyl,

3-[(tetrazolium-5-base is amino) carbonyl] phenyl,

The 3-[[(trifluoromethyl) alkylsulfonyl] amino] phenyl,

3-(tetrazolium-5-yl) phenyl,

4-fluoro-3-(tetrazolium-5-yl) phenyl,

2-fluoro-5-(tetrazolium-5-yl) phenyl,

3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl,

4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl,

3-(5-oxo-4,5-dihydro-1,3,4-oxadiazole-2-yl) phenyl,

3-(5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl) phenyl,

3-(1,3,4-oxadiazole-2-yl) phenyl,

3-(1,2,4-triazole-3-yl) phenyl,

3-[5-(trifluoromethyl)-1,2,4-triazole-3-yl] phenyl,

3-(5-oxyethyl group-1,2,4-triazole-3-yl) phenyl,

Pyridin-3-yl,

6-fluoro-pyridin-3-yl,

6-methoxypyridine-3-base,

6-oxo-1,6-dihydropyridine-3-base,

1-methyl-6-oxo-1,6-dihydropyridine-3-base,

1-ethyl-6-oxo-1,6-dihydropyridine-3-base,

5-bromo-1-methyl-6-oxo-1,6-dihydropyridine-3-base,

Imidazo [1,2-a] pyridine-6-base,

[1,2,4] triazolo [4,3-a] pyridine-6-base,

3-(trifluoromethyl) [1,2,4] triazolo [4,3-a] pyridine-6-base,

3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base,

2-methyl-3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base,

4-amido quinazoline-6-base,

2-(kharophen) imidazo [1,2-a] pyridine-6-base,

The 3-aminooimidazole is [1,2-a] pyridine-6-base also,

3-carboxyl pyrazolo [1,5-a] pyridine-5-base,

5-bromo-6-oxo-1,6-dihydropyridine-3-base,

[1,2,4] triazolo [1,5-a] pyridine-6-base,

[1,2,4] triazolo [1,5-a] pyridine-7-base,

Pyrazolo [1,5-a] pyrimidine-5-base.

17. the compound of the structural formula Ij of claim 14:

R wherein ²Be selected from:

Methyl,

Ethyl,

CH ₂-cyclopropyl,

COOH，

COOMe，

COOEt，

CONHMe，

CONMe ₂，

CONH ₂，

CONHEt，

CONMeCH ₂Ph，

Tetramethyleneimine-1-base carbonyl,

Azetidine-1-base carbonyl,

3-fluorine azetidine-1-base carbonyl,

Morpholine-4-base carbonyl,

[(tetrazolium-5-yl) amino] carbonyl;

R ⁴Be selected from:

The 4-fluorophenyl,

The 2,4 difluorobenzene base,

3, the 4-difluorophenyl,

The 2-chloro-phenyl-,

The 2-fluorophenyl,

3-(methyl sulphonyl) phenyl,

3-(ethoxy carbonyl) phenyl,

The 3-carboxyl phenyl,

3-(aminocarboxyl) phenyl,

3-[(tertiary butyl amino) carbonyl] phenyl,

The 3-[(phenyl amino) carbonyl] phenyl,

3-[(thiazol-2-yl amino) carbonyl] phenyl,

3-[(tetrazolium-5-base is amino) carbonyl] phenyl,

The 3-[[(trifluoromethyl) alkylsulfonyl] amino] phenyl,

3-(tetrazolium-5-yl) phenyl,

4-fluoro-3-(tetrazolium-5-yl) phenyl,

2-fluoro-5-(tetrazolium-5-yl) phenyl,

3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl,

4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl,

3-(5-oxo-4,5-dihydro-1,3,4-oxadiazole-2-yl) phenyl,

3-(5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl) phenyl,

3-(1,3,4-oxadiazole-2-yl) phenyl,

3-(1,2,4-triazole-3-yl) phenyl,

3-[5-(trifluoromethyl)-1,2,4-triazole-3-yl] phenyl,

3-(5-oxyethyl group-1,2,4-triazole-3-yl) phenyl,

Pyridin-3-yl,

6-fluoro-pyridin-3-yl,

6-methoxypyridine-3-base,

6-oxo-1,6-dihydropyridine-3-base,

1-methyl-6-oxo-1,6-dihydropyridine-3-base,

1-ethyl-6-oxo-1,6-dihydropyridine-3-base,

5-bromo-1-methyl-6-oxo-1,6-dihydropyridine-3-base,

Imidazo [1,2-a] pyridine-6-base,

[1,2,4] triazolo [4,3-a] pyridine-6-base,

3-(trifluoromethyl) [1,2,4] triazolo [4,3-a] pyridine-6-base,

3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base,

2-methyl-3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base,

4-amido quinazoline-6-base,

2-(kharophen) imidazo [1,2-a] pyridine-6-base,

The 3-aminooimidazole is [1,2-a] pyridine-6-base also,

3-carboxyl pyrazolo [1,5-a] pyridine-5-base,

5-bromo-6-oxo-1,6-dihydropyridine-3-base,

[1,2,4] triazolo [1,5-a] pyridine-6-base,

[1,2,4] triazolo [1,5-a] pyridine-7-base,

Pyrazolo [1,5-a] pyrimidine-5-base.

18. the compound of claim 17, wherein W is CH ₂, X is CHF or CF ₂

19. the compound of claim 17 or its pharmacy acceptable salt, the structural formula of described compound is selected from:

R ⁴ R ² X Phenyl Me (S)-CHF 4-(SO ₂Me)-phenyl Me (S)-CHF 3-(SO ₂Me)-phenyl Me (S)-CHF Pyrazine-5-base Me (S)-CHF 3-chloropyridine-4-base Me (S)-CHF The 2,4 difluorobenzene base Me (S)-CHF 3, the 4-difluorophenyl Me (S)-CHF 2, the 5-difluorophenyl Me (S)-CHF

3, the 5-difluorophenyl Me (S)-CHF 3-(ethoxy carbonyl) phenyl Me (S)-CHF 4-(ethoxy carbonyl) phenyl Me (S)-CHF 3-(NHSO ₂Me)-phenyl Me (S)-CHF 4-(NHSO ₂Me)-phenyl Me (S)-CHF 4-CO ₂The H-phenyl Me (S)-CHF Pyridin-3-yl Me (S)-CHF The 6-OMe-pyridin-3-yl Me (S)-CHF The 2-Cl-phenyl Me (S)-CHF The 2-F-phenyl Me (S)-CHF The 3-CN-phenyl Me (S)-CHF Pyridin-4-yl Me (S)-CHF Pyridine-2-base Me (S)-CHF The 2,4 difluorobenzene base The cyclopropyl methyl (S)-CHF 3-(MeSO ₂)-phenyl The cyclopropyl methyl (S)-CHF 4-fluoro-(3-tetrazolium-5-yl) phenyl Me (S)-CHF 3-(amino-sulfonyl) phenyl Me (S)-CHF 2-fluoro-(5-tetrazolium-5-yl) phenyl Me (S)-CHF The 3-hydroxy phenyl Me (S)-CHF 6-fluorine pyridin-3-yl Me (S)-CHF 3-(aminocarboxyl) phenyl Me (S)-CHF 3-(phenyl amino carbonyl) phenyl Me (S)-CHF 4-fluoro-3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl Me (S)-CHF The 3-[(thiazol-2-yl) aminocarboxyl] phenyl Me (S)-CHF 3-[(tetrazolium-5-yl) aminocarboxyl] phenyl Me (S)-CHF Imidazo [1,2-a] pyridine-6-base Me (S)-CHF

The 2-p-methoxy-phenyl Me (S)-CHF 3-(5-oxo-4,5-dihydro-1,3,4-oxadiazole-2-yl) phenyl Me (S)-CHF 3-(5-oxyethyl group-1H-1,2,4-triazole-3-yl) phenyl Me (S)-CHF 1-ethyl-6-oxo-1,6-dihydropyridine-3-base Me (S)-CHF Quinoline-6-base Me (S)-CHF 3-(2-oxo-2,3-dihydro-1H-imidazol-4 yl) phenyl Me (S)-CHF The 2-aminomethyl phenyl Me (S)-CHF 2-(trifluoromethyl) phenyl Me (S)-CHF 3-[5-(trifluoromethyl)-4H-1,2,4-triazole-3-yl] phenyl Me (S)-CHF 4-oxo-3,4-dihydroquinazoline-6-base Me (S)-CHF The 4-fluorophenyl CONHEt (S)-CHF 1-methyl-6-oxo-1,6-dihydropyridine-3-base Et (S)-CHF 6-oxo-1,6-dihydropyridine-3-base Et (S)-CHF 3-(trifluoromethyl) [1,2,4] triazolo [4,3-a] pyridine-6-base Me (S)-CHF The 4-fluorophenyl CONH ₂ (S)-CHF 3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base Me (S)-CHF 1-methyl-6-oxo-1,6-dihydropyridine-3-base Me CF ₂ 6-oxo-1,6-dihydropyridine-3-base Me CF ₂ 4-amido quinazoline-6-base Me (S)-CHF 5-bromo-1-methyl-6-oxo-1,6-dihydropyridine-3-base Me (S)-CHF

The 4-fluorophenyl (tetramethyleneimine-1-yl) carbonyl (S)-CHF The 4-fluorophenyl (azetidine-1-yl) carbonyl (S)-CHF 2-(acetylamino) imidazo [1,2-a] pyridine-6-base Me (S)-CHF 2-methyl-3-oxo-2,3-dihydro [1,2,4] triazolo [4,3-a] pyridine-6-base Me (S)-CHF The 3-aminooimidazole is [1,2-a] pyridine-6-base also Me (S)-CHF The 4-fluorophenyl [(tetrazolium-5-yl) amino] carbonyl (S)-CHF The 4-fluorophenyl CONHMe (S)-CHF The 4-fluorophenyl CONEt ₂ (S)-CHF The 4-fluorophenyl COOMe (S)-CHF [1,2,4] triazolo [1,5-a] pyridine-6-base Me (S)-CHF The 4-fluorophenyl COOH CH ₂ The 4-fluorophenyl COOH CF ₂ The 4-fluorophenyl CONMe ₂ CH ₂ 3-carboxyl pyrazolo [1,5-a] pyridine-5-base Me (S)-CHF 6-oxo-1,6-dihydropyridine-3-base Me CH ₂ The 4-fluorophenyl Me (S)-CHF The 3-carboxyl phenyl Me (S)-CHF 3-(tetrazolium-5-yl) phenyl Me (S)-CHF 3-(5-oxo-4,5-dihydro-1,2,4-oxadiazole-3-yl) phenyl Me (S)-CHF 6-oxo-1,6-dihydropyridine-3-base Me (S)-CHF 1-methyl-6-oxo-1,6-dihydropyridine-3-base The cyclopropyl methyl (S)-CHF 5-bromo-6-oxo-1,6-dihydropyridine-3-base Me (S)-CHF

3-[(tertiary butyl amino) carbonyl] phenyl Me (S)-CHF The 3-[[(trifluoromethyl) alkylsulfonyl] amino] phenyl Me (S)-CHF [1,2,4] triazolo [4,3-a] pyridine-6-base Me (S)-CHF The 4-fluorophenyl COOH (S)-CHF The 4-fluorophenyl CONMe ₂ (S)-CHF [1,2,4] triazolo [1,5-a] pyridine-6-base CONMe ₂ (S)-CHF [1,2,4] triazolo [1,5-a] pyridine-7-base CONMe ₂ (S)-CHF Pyrazolo [1,5-a] pyrimidine-5-base CONMe ₂ (S)-CHF [1,2,4] triazolo [1,5-a] pyridine-6-base CONMe ₂ CF ₂ [1,2,4] triazolo [1,5-a] pyridine-7-base CONMe ₂ CF ₂ Pyrazolo [1,5-a] pyrimidine-5-base CONMe ₂ CF ₂ [1,2,4] triazolo [1,5-a] pyridine-6-base CONMe ₂ CH ₂ [1,2,4] triazolo [1,5-a] pyridine-6-base (3-fluoro-azetidine-1-yl) carbonyl CH ₂ [1,2,4] triazolo [1,5-a] pyridine-6-base (tetramethyleneimine-1-yl) carbonyl CF ₂ [1,2,4] triazolo [1,5-a] pyridine-6-base CONMeCH ₂Ph CF ₂ [1,2,4] triazolo [1,5-a] pyridine-6-base (morpholine-4-yl) carbonyl CF ₂

20. the compound of claim 16 or its pharmacy acceptable salt, the structural formula of described compound is selected from:

R ⁴ R ² X The 4-fluorophenyl Me CHF 1-methyl-6-oxo-1,6-dihydropyridine-3-base Me CHF

[1,2,4] triazolo [4,3-a] pyridine-6-base Me CHF Imidazo [1,2-a] pyridine-6-base Me CHF The 4-fluorophenyl COOH CHF The 4-fluorophenyl CONMe ₂ CHF [1,2,4] triazolo [1,5-a] pyridine-6-base CONMe ₂ CHF

21. the compound of claim 16 or its pharmacy acceptable salt, the structural formula of described compound is selected from:

R ⁴ R ² X The 4-fluorophenyl Me CHF 1-methyl-6-oxo-1,6-dihydropyridine-3-base Me CHF [1,2,4] triazolo [4,3-a] pyridine-6-base Me CHF Imidazo [1,2-a] pyridine-6-base Me CHF

22. the compound of claim 16 or its pharmacy acceptable salt, described compound is selected from:

23. the compound of claim 22 or its pharmacy acceptable salt, described compound is

24. the compound of claim 22 or its pharmacy acceptable salt, described compound is

25. the compound of claim 22 or its pharmacy acceptable salt, described compound is

26. the compound of claim 22 or its pharmacy acceptable salt, described compound is

27. the compound of claim 22 or its pharmacy acceptable salt, described compound is

28. a pharmaceutical composition, described pharmaceutical composition comprise the compound and the pharmaceutically acceptable carrier of claim 1.

29. the method for treatment diabetes in the Mammals that needs is arranged, described method comprise the compound of the claim 1 that gives Mammals treatment significant quantity.

30. a method for the treatment of non-insulin-dependent (2 type) diabetes in the Mammals that needs is arranged, described method comprise the compound of the claim 1 that gives Mammals treatment significant quantity.

31. the method for a treatment hyperglycemia in the Mammals that needs is arranged, described method comprise the compound of the claim 1 that gives Mammals treatment significant quantity.

32. the method for a treatment of obesity in the Mammals that needs is arranged, described method comprise the compound of the claim 1 that gives Mammals treatment significant quantity.

33. the method for treatment diabetes in the Mammals that needs is arranged, described method comprise compound and the PPAR α/γ dual agonists KRP-297 that unites the claim 1 that gives Mammals treatment significant quantity.

34. the method for treatment diabetes in the Mammals that needs is arranged, described method comprise the compound and the N1,N1-Dimethylbiguanide of uniting the claim 1 that gives Mammals treatment significant quantity.